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Plasma membranes enclose and define the borders between the inside and the outside of cells. In addition, the plasma membrane must, in some cases, be flexible enough to allow certain cells, such as amoebae, to change shape and direction as they move through the environment, hunting smaller, single-celled organisms.
A subgoal in our "build-a-cell" design challenge is to create a boundary that separates the "inside" of the cell from the environment "outside". This boundary needs to serve multiple functions that include:
- Act as a barrier by blocking some compounds from moving in and out of the cell.
- Be selectively permeable in order to transport specific compounds into and out of the cell.
- Receive, sense, and transmit signals from the environment to inside of the cell.
- Project "self" to others by communicating identity to other nearby cells.
Figure 1. The diameter of a typical balloon is 25cm and the thickness of the plastic of the balloon of around 0.25mm. This is a 1000X difference. A typical eukaryotic cell will have a cell diameter of about 50µm and a cell membrane thickness of 5nm. This is a 10,000X difference.
Note: possible discussion
The ratio of membrane thickness compared to the size of an average eukaryotic cell is much greater compared to that of a balloon stretched with air. To think that the boundary between life and nonlife is so small, and seemingly fragile, more so than a balloon, suggests that structurally the membrane must be relatively stable. Discuss why cellular membranes are stable. You will need to pull from information we have already covered in this class.
Fluid mosaic model
The existence of the plasma membrane was identified in the 1890s, and its chemical components were identified in 1915. The principal components identified at that time were lipids and proteins. The first widely accepted model of the plasma membrane’s structure was proposed in 1935 by Hugh Davson and James Danielli; it was based on the “railroad track” appearance of the plasma membrane in early electron micrographs. They theorized that the structure of the plasma membrane resembles a sandwich, with protein being analogous to the bread, and lipids being analogous to the filling. In the 1950s, advances in microscopy, notably transmission electron microscopy (TEM), allowed researchers to see that the core of the plasma membrane consisted of a double, rather than a single, layer. A new model that better explains both the microscopic observations and the function of that plasma membrane was proposed by S.J. Singer and Garth L. Nicolson in 1972.
The explanation proposed by Singer and Nicolson is called the fluid mosaic model. The model has evolved somewhat over time, but it still best accounts for the structure and functions of the plasma membrane as we now understand them. The fluid mosaic model describes the structure of the plasma membrane as a mosaic of components—including phospholipids, cholesterol, proteins, and carbohydrates—that gives the membrane a fluid character. Plasma membranes range from 5 to 10 nm in thickness. For comparison, human red blood cells, visible via light microscopy, are approximately 8 µm wide, or approximately 1,000 times wider than a plasma membrane.
Figure 2. The fluid mosaic model of the plasma membrane describes the plasma membrane as a fluid combination of phospholipids, cholesterol, and proteins. Carbohydrates attached to lipids (glycolipids) and to proteins (glycoproteins) extend from the outward-facing surface of the membrane.
The principal components of a plasma membrane are lipids (phospholipids and cholesterol), proteins, and carbohydrates. The proportions of proteins, lipids, and carbohydrates in the plasma membrane vary with organism and cell type, but for a typical human cell, proteins account for about 50 percent of the composition by mass, lipids (of all types) account for about 40 percent of the composition by mass, and carbohydrates account for the remaining 10 percent of the composition by mass. However, the concentration of proteins and lipids varies with different cell membranes. For example, myelin, an outgrowth of the membrane of specialized cells, insulates the axons of the peripheral nerves, contains only 18 percent protein and 76 percent lipid. The mitochondrial inner membrane contains 76 percent protein and only 24 percent lipid. The plasma membrane of human red blood cells is 30 percent lipid. Carbohydrates are present only on the exterior surface of the plasma membrane and are attached to proteins, forming glycoproteins, or attached to lipids, forming glycolipids.
Phospholipids are major constituents of the cell membrane, the outermost layer of cells. Like fats, they are composed of fatty acid chains attached to a polar head group. Specifically, there are two fatty acid tails and a phosphate group as the polar head group. The phospholipid is an amphipathic molecule, meaning it has a hydrophobic part and a hydrophilic part. The fatty acid chains are hydrophobic and cannot interact with water, whereas the phosphate-containing head group is hydrophilic and interacts with water.
Make sure to note in Figure 3 that the phosphate group has an R group linked to one of the oxygen atoms. R is a variable commonly used in these types of diagrams to indicate that some other atom or molecule is bound at that position. That part of the molecule can be different in different phospholipids—and will impart some different chemistry to the whole molecule. At the moment, however, you are responsible for being able to recognize this type of molecule (no matter what the R group is) because of the common core elements—the glycerol backbone, the phosphate group, and the two hydrocarbon tails.
Figure 3. A phospholipid is a molecule with two fatty acids and a modified phosphate group attached to a glycerol backbone. The phosphate may be modified by the addition of charged or polar chemical groups. Several chemical R groups may modify the phosphate. Choline, serine, and ethanolamine are shown here. These attach to the phosphate group at the position labeled R via their hydroxyl groups.
Attribution: Marc T. Facciotti (own work)
A phospholipid bilayer forms as the basic structure of the cell membrane. The fatty acid tails of phospholipids face inside, away from water, whereas the phosphate group faces outside, hydrogen bonding with water. Phospholipids are responsible for the dynamic nature of the plasma membrane. The phospholipids will spontaneously form a structure known as a micelle in which the hydrophilic phosphate heads face the outside and the fatty acids face the interior of this structure.
Figure 4. In the presence of water, some phospholipids will spontaneously arrange themselves into a micelle. The lipids will be arranged such that their polar groups will be on the outside of the micelle, and the nonpolar tails will be on the inside. A lipid bilayer can also form, a two layered sheet only a few nanometers thick. The lipid bilayer consists of two layers of phospholipids organized in a way that all the hydrophobic tails align side by side in the center of the bilayer and are surrounded by the hydrophilic head groups.
Source: Created by Erin Easlon (own work)
Note: possible discussion
Above it says that if you were to take some pure phospholipids and drop them into water that some if it would spontaneously (on its own) form into micelles. This sounds a lot like something that could be described by an energy story. Go back to the energy story rubric and try to start creating an energy story for this process—I expect that the steps involving the description of energy might be difficult at this point (we'll come back to that later) but you should be able to do at least the first three steps. You can constructively critique (politely) each other's work to create an optimized story.
Note: possible discussion
Note that the phospholipid depicted above has an R group linked to the phosphate group. Recall that this designation is generic—these can be different than the R groups on amino acids. What might be a benefit/purpose of "functionalizing" or "decorating" different lipids with different R groups? Think of the functional requirements for membranes stipulated above.
Proteins make up the second major component of plasma membranes. Integral proteins (some specialized types are called integrins) are, as their name suggests, integrated completely into the membrane structure, and their hydrophobic membrane-spanning regions interact with the hydrophobic region of the the phospholipid bilayer. Single-pass integral membrane proteins usually have a hydrophobic transmembrane segment that consists of 20–25 amino acids. Some span only part of the membrane—associating with a single layer—while others stretch from one side of the membrane to the other, and are exposed on either side. Some complex proteins are composed of up to 12 segments of a single protein, which are extensively folded and embedded in the membrane. This type of protein has a hydrophilic region or regions, and one or several mildly hydrophobic regions. This arrangement of regions of the protein tends to orient the protein alongside the phospholipids, with the hydrophobic region of the protein adjacent to the tails of the phospholipids and the hydrophilic region or regions of the protein protruding from the membrane and in contact with the cytosol or extracellular fluid. Peripheral proteins are found on either the exterior or interior surfaces of membranes; and weakly or temporarily associated with the membranes. They can be attached (interact with) either to integral membrane proteins or simply interact weakly with the phospholipids within the membrane.
Figure 5. Integral membranes proteins may have one or more α-helices (pink cylinders) that span the membrane (examples 1 and 2), or they may have β-sheets (blue rectangles) that span the membrane (example 3). (credit: “Foobar”/Wikimedia Commons)
Carbohydrates are the third major component of plasma membranes. They are always found on the exterior surface of cells and are bound either to proteins (forming glycoproteins) or to lipids (forming glycolipids). These carbohydrate chains may consist of 2–60 monosaccharide units and can be either straight or branched. Along with peripheral proteins, carbohydrates form specialized sites on the cell surface that allow cells to recognize each other (one of the core functional requirements noted above in "cellular membranes").
The mosaic characteristic of the membrane, described in the fluid mosaic model, helps to illustrate its nature. The integral proteins and lipids exist in the membrane as separate molecules and they "float" in the membrane, moving somewhat with respect to one another. The membrane is not like a balloon, however, that can expand and contract; rather, it is fairly rigid and can burst if penetrated or if a cell takes in too much water. However, because of its mosaic nature, a very fine needle can easily penetrate a plasma membrane without causing it to burst, and the membrane will flow and self-seal when the needle is extracted.
The mosaic characteristics of the membrane explain some but not all of its fluidity. There are two other factors that help maintain this fluid characteristic. One factor is the nature of the phospholipids themselves. In their saturated form, the fatty acids in phospholipid tails are saturated with bound hydrogen atoms. There are no double bonds between adjacent carbon atoms. This results in tails that are relatively straight. In contrast, unsaturated fatty acids do not contain a maximal number of hydrogen atoms, but they do contain some double bonds between adjacent carbon atoms; a double bond results in a bend in the string of carbons of approximately 30 degrees.
Figure 6. Any given cell membrane will be composed of a combination of saturated and unsaturated phospholipids. The ratio of the two will influence the permeability and fluidity of the membrane. A membrane composed of completely saturated lipids will be dense and less fluid, and a membrane composed of completely unsaturated lipids will be very loose and very fluid.
Note: possible discussion
Organisms can be found living in extreme temperature conditions. Both in extreme cold or extreme heat. What types of differences would you expect to see in the lipid composition of organisms that live at these extremes?
Saturated fatty acids, with straight tails, are compressed by decreasing temperatures, and they will press in on each other, making a dense and fairly rigid membrane. When unsaturated fatty acids are compressed, the “kinked” tails elbow adjacent phospholipid molecules away, maintaining some space between the phospholipid molecules. This “elbow room” helps to maintain fluidity in the membrane at temperatures at which membranes with high concentrations of saturated fatty acid tails would “freeze” or solidify. The relative fluidity of the membrane is particularly important in a cold environment. A cold environment tends to compress membranes composed largely of saturated fatty acids, making them less fluid and more susceptible to rupturing. Many organisms (fish are one example) are capable of adapting to cold environments by changing the proportion of unsaturated fatty acids in their membranes in response to the lowering of the temperature.
Animals have an additional membrane constituent that assists in maintaining fluidity. Cholesterol, which lies alongside the phospholipids in the membrane, tends to dampen the effects of temperature on the membrane. Thus, this lipid functions as a fluidity buffer, preventing lower temperatures from inhibiting fluidity and preventing increased temperatures from increasing fluidity too much. Thus, cholesterol extends, in both directions, the range of temperature in which the membrane is appropriately fluid and consequently functional. Cholesterol also serves other functions, such as organizing clusters of transmembrane proteins into lipid rafts.
Figure 7. Cholesterol fits between the phospholipid groups within the membrane.
Review of the components of the membrane
The components and functions of the plasma membrane
Main fabric of the membrane
Between phospholipids and between the two phospholipid layers of animal cells
Integral proteins (e.g., integrins)
Embedded within the phospholipid layer(s); may or may not penetrate through both layers
On the inner or outer surface of the phospholipid bilayer; not embedded within the phospholipids
Carbohydrates (components of glycoproteins and glycolipids)
|Generally attached to proteins on the outside membrane layer|
The chemistry of living things occurs in aqueous solutions, and balancing the concentrations of those solutions is an ongoing problem. In living systems, diffusion of substances into and out of cells is mediated by the plasma membrane. The passive forms of transport, diffusion and osmosis, move nonpolar materials of small molecular weight across membranes. Substances diffuse from areas of high concentration to areas of lower concentration, and this process continues until the substance is evenly distributed in a system. In solutions containing more than one substance, each type of molecule diffuses according to its own concentration gradient, independent of the diffusion of other substances. Some materials diffuse readily through the membrane, but others are hindered, and their passage is made possible by specialized proteins, such as channels and transporters.
Transport across the membrane
One of the great wonders of the cell membrane is its ability to regulate the concentration of substances inside the cell. These substances include ions such as Ca2+, Na+, K+, and Cl–; nutrients including sugars, fatty acids, and amino acids; and waste products, particularly carbon dioxide (CO2), which must leave the cell.
Design Challenge Subproblem:
Controlling what enters and exits the cell.
The membrane’s lipid bilayer structure provides the first level of control. The phospholipids are tightly packed together, and the membrane has a hydrophobic interior. This structure causes the membrane to be selectively permeable. A membrane that has selective permeability allows only substances meeting certain criteria to pass through it unaided. In the case of the cell membrane, only relatively small, nonpolar materials can move through the lipid bilayer at biologically relevant rates (remember, the lipid tails of the membrane are nonpolar). The rates of transport of various molecules is tabulated in the Membranes section. All substances that move through the membrane do so by one of two general methods, which are categorized based on whether or not the transport process is exergonic or endergonic. Passive transport is the exergonic movement of substances across the membrane. In contrast, active transport is the endergonic movement of substances across the membrane that is coupled to an exergonic reaction.
Passive transport does not require the cell to expend energy. In passive transport, substances move from an area of higher concentration to an area of lower concentration, down the concentration gradient and energetically favorable. Depending on the chemical nature of the substance, different processes may be associated with passive transport.
Diffusion is a passive process of transport. A single substance tends to move from an area of high concentration to an area of low concentration until the concentration is equal across a space. You are familiar with diffusion of substances through the air. For example, think about someone opening a bottle of ammonia in a room filled with people. The ammonia gas is at its highest concentration in the bottle; its lowest concentration is at the edges of the room. The ammonia vapor will diffuse, or spread away, from the bottle, and gradually, more and more people will smell the ammonia as it spreads. Materials move within the cell’s cytosol by diffusion, and certain materials move through the plasma membrane by diffusion.
Figure 1. Diffusion through a permeable membrane moves a substance from an area of high concentration (extracellular fluid, in this case) down its concentration gradient (into the cytoplasm). Each separate substance in a medium, such as the extracellular fluid, has its own concentration gradient, independent of the concentration gradients of other materials. In addition, each substance will diffuse according to that gradient. Within a system, there will be different rates of diffusion of the different substances in the medium.
(Attribution: Mariana Ruiz Villareal, modified)
Factors That Affect Diffusion
If unconstrained, molecules will move through and explore space randomly at a rate that depends on their size, their shape, their environment, and their thermal energy. This type of movement underlies the diffusive movement of molecules through whatever medium they are in. The absence of a concentration gradient does not mean that this movement will stop, just that there may be no net movement of the number of molecules from one area to another, a condition known as dynamic equilibrium.
Factors influencing diffusion include:
- Extent of the concentration gradient: The greater the difference in concentration, the more rapid the diffusion. The closer the distribution of the material gets to equilibrium, the slower the rate of diffusion becomes.
- Shape, size and mass of the molecules diffusing: Large and heavier molecules move more slowly; therefore, they diffuse more slowly. The reverse is typically true for smaller, lighter molecules.
- Temperature: Higher temperatures increase the energy and therefore the movement of the molecules, increasing the rate of diffusion. Lower temperatures decrease the energy of the molecules, thus decreasing the rate of diffusion.
- Solvent density: As the density of a solvent increases, the rate of diffusion decreases. The molecules slow down because they have a more difficult time getting through the denser medium. If the medium is less dense, rates of diffusion increase. Since cells primarily use diffusion to move materials within the cytoplasm, any increase in the cytoplasm’s density will decrease the rate at which materials move in the cytoplasm.
- Solubility: As discussed earlier, nonpolar or lipid-soluble materials pass through plasma membranes more easily than polar materials, allowing a faster rate of diffusion.
- Surface area and thickness of the plasma membrane: Increased surface area increases the rate of diffusion, whereas a thicker membrane reduces it.
- Distance travelled: The greater the distance that a substance must travel, the slower the rate of diffusion. This places an upper limitation on cell size. A large, spherical cell will die because nutrients or waste cannot reach or leave the center of the cell, respectively. Therefore, cells must either be small in size, as in the case of many prokaryotes, or be flattened, as with many single-celled eukaryotes.
In facilitated transport, also called facilitated diffusion, materials diffuse across the plasma membrane with the help of membrane proteins. A concentration gradient exists that allows these materials to diffuse into or out of the cell without expending cellular energy. In the case that the materials are ions or polar molecules, compounds that are repelled by the hydrophobic parts of the cell membrane, facilitated transport proteins help shield these materials from the repulsive force of the membrane, allowing them to diffuse into the cell.
Note: Possible Discussion
Compare and contrast passive diffusion and facilitated diffusion.
The integral proteins involved in facilitated transport are collectively referred to as transport proteins, and they function as either channels for the material or carriers. In both cases, they are transmembrane proteins. Different channel proteins have different transport properties. Some have evolved to be have very high specificity for the substance that is being transported while others transport a variety of molecules sharing some common characteristic(s). The interior "passageway" of channel proteins have evolved to provide a low energetic barrier for transport of substances across the membrane through the complementary arrangement of amino acid functional groups (of both backbone and side-chains). Passage through the channel allows polar compounds to avoid the nonpolar central layer of the plasma membrane that would otherwise slow or prevent their entry into the cell. While at any one time significant amounts of water crosses the membrane both in and out the rate of individual water molecule transport may not be fast enough to adapt to changing environmental conditions. For such cases Nature has evolved a special class of membrane proteins called aquaporins that allow water to pass through the membrane at a very high rate.
Figure 2: Facilitated transport moves substances down their concentration gradients. They may cross the plasma membrane with the aid of channel proteins.
(Attribution: Mariana Ruiz Villareal, modified.)
Channel proteins are either open at all times or they are “gated.” The latter controls the opening of the channel. Various mechanisms may be involved in the gating mechanism. For instance, the attachment of a specific ion or small molecule to the channel protein may trigger opening. Changes in local membrane "stress" or changes in voltage across the membrane may also be triggers to open or close a channel.
Different organisms and tissues in multicellular species express different sets of channel proteins in their membranes depending on the environments they live in or specialized function they play in an organisms. This provides each type of cell with a unique membrane permeability profile that is evolved to complement its "needs" (note the anthropomorphism). For example, in some tissues, sodium and chloride ions pass freely through open channels, whereas in other tissues a gate must be opened to allow passage. This occurs in the kidney, where both forms of channels are found in different parts of the renal tubules. Cells involved in the transmission of electrical impulses, such as nerve and muscle cells, have gated channels for sodium, potassium, and calcium in their membranes. Opening and closing of these channels changes the relative concentrations on opposing sides of the membrane of these ions, resulting a change in electrical potential across the membrane that lead to message propagation in the case of nerve cells or in muscle contraction in the case of muscle cells.
Another type of protein embedded in the plasma membrane is a carrier protein. This aptly named protein binds a substance and, in doing so, triggers a change of its own shape, moving the bound molecule from the outside of the cell to its interior; depending on the gradient, the material may move in the opposite direction. Carrier proteins are typically specific for a single substance. This selectivity adds to the overall selectivity of the plasma membrane. The molecular-scale mechanism of function for these proteins remains poorly understood.
Carrier proteins play an important role in the function of kidneys. Glucose, water, salts, ions, and amino acids needed by the body are filtered in one part of the kidney. This filtrate, which includes glucose, is then reabsorbed in another part of the kidney with the help of carrier proteins. Because there are only a finite number of carrier proteins for glucose, if more glucose is present in the filtrate than the proteins can handle, the excess is not reabsorbed and it is excreted from the body in the urine. In a diabetic individual, this is described as “spilling glucose into the urine.” A different group of carrier proteins called glucose transport proteins, or GLUTs, are involved in transporting glucose and other hexose sugars through plasma membranes within the body.
Channel and carrier proteins transport materials at different rates. Channel proteins transport much more quickly than do carrier proteins. Channel proteins facilitate diffusion at a rate of tens of millions of molecules per second, whereas carrier proteins work at a rate of a thousand to a million molecules per second.
Note: A note of appreciation
The rates of transport just discussed are astounding. Recall that these molecular catalysts are on the scale of 10s of nanometers (10-9 meters) and that they are composed of a self-folding string of 20 amino acids and the relatively small selection of chemical functional groups that they carry.
Osmosis is the movement of water through a semipermeable membrane according to the concentration gradient of water across the membrane, which is inversely proportional to the concentration of solutes. While diffusion transports material across membranes and within cells, osmosis transports only water across a membrane and the membrane limits the diffusion of solutes in the water. Not surprisingly, the aquaporins that facilitate water movement play a large role in osmosis, most prominently in red blood cells and the membranes of kidney tubules.
Osmosis is a special case of diffusion. Water, like other substances, moves from an area of high concentration to one of low concentration. An obvious question is what makes water move at all? Imagine a beaker with a semipermeable membrane separating the two sides or halves. On both sides of the membrane the water level is the same, but there are different concentrations of a dissolved substance, or solute, that cannot cross the membrane (otherwise the concentrations on each side would be balanced by the solute crossing the membrane). If the volume of the solution on both sides of the membrane is the same, but the concentrations of solute are different, then there are different amounts of water, the solvent, on either side of the membrane.
To illustrate this, imagine two full glasses of water. One has a single teaspoon of sugar in it, whereas the second one contains one-quarter cup of sugar. If the total volume of the solutions in both cups is the same, which cup contains more water? Because the large amount of sugar in the second cup takes up much more space than the teaspoon of sugar in the first cup, the first cup has more water in it.
Returning to the beaker example, recall that it has a mixture of solutes on either side of the membrane. A principle of diffusion is that the molecules move around and will spread evenly throughout the medium if they can. However, only the material capable of getting through the membrane will diffuse through it. In this example, the solute cannot diffuse through the membrane, but the water can. Water has a concentration gradient in this system. Thus, water will diffuse down its concentration gradient, crossing the membrane to the side where it is less concentrated. This diffusion of water through the membrane—osmosis—will continue until the concentration gradient of water goes to zero or until the hydrostatic pressure of the water balances the osmotic pressure. Osmosis proceeds constantly in living systems.
Tonicity describes how an extracellular solution can change the volume of a cell by affecting osmosis. A solution's tonicity often directly correlates with the osmolarity of the solution. Osmolarity describes the total solute concentration of the solution. A solution with low osmolarity has a greater number of water molecules relative to the number of solute particles; a solution with high osmolarity has fewer water molecules with respect to solute particles. In a situation in which solutions of two different osmolarities are separated by a membrane permeable to water, though not to the solute, water will move from the side of the membrane with lower osmolarity (and more water) to the side with higher osmolarity (and less water). This effect makes sense if you remember that the solute cannot move across the membrane, and thus the only component in the system that can move—the water—moves along its own concentration gradient. An important distinction that concerns living systems is that osmolarity measures the number of particles (which may be molecules) in a solution. Therefore, a solution that is cloudy with cells may have a lower osmolarity than a solution that is clear, if the second solution contains more dissolved molecules than there are cells.
Three terms—hypotonic, isotonic, and hypertonic—are used to relate the osmolarity of a cell to the osmolarity of the extracellular fluid that contains the cells. In a hypotonicsituation, the extracellular fluid has lower osmolarity than the fluid inside the cell, and water enters the cell. (In living systems, the point of reference is always the cytoplasm, so the prefix hypo- means that the extracellular fluid has a lower concentration of solutes, or a lower osmolarity, than the cell cytoplasm.) It also means that the extracellular fluid has a higher concentration of water in the solution than does the cell. In this situation, water will follow its concentration gradient and enter the cell.
As for a hypertonic solution, the prefix hyper- refers to the extracellular fluid having a higher osmolarity than the cell’s cytoplasm; therefore, the fluid contains less water than the cell does. Because the cell has a relatively higher concentration of water, water will leave the cell.
In an isotonic solution, the extracellular fluid has the same osmolarity as the cell. If the osmolarity of the cell matches that of the extracellular fluid, there will be no net movement of water into or out of the cell, although water will still move in and out. Blood cells and plant cells in hypertonic, isotonic, and hypotonic solutions take on characteristic appearances.
A doctor injects a patient with what the doctor thinks is an isotonic saline solution. The patient dies, and an autopsy reveals that many red blood cells have been destroyed. Do you think the solution the doctor injected was really isotonic?
Link to Learning:
For a video illustrating the process of diffusion in solutions, visit this site.
Tonicity in Living Systems
In a hypotonic environment, water enters a cell, and the cell swells. In an isotonic condition, the relative concentrations of solute and solvent are equal on both sides of the membrane. There is no net water movement; therefore, there is no change in the size of the cell. In a hypertonic solution, water leaves a cell and the cell shrinks. If either the hypo- or hyper- condition goes to excess, the cell’s functions become compromised, and the cell may be destroyed.
A red blood cell will burst, or lyse, when it swells beyond the plasma membrane’s capability to expand. Remember, the membrane resembles a mosaic, with discrete spaces between the molecules composing it. If the cell swells, and the spaces between the lipids and proteins become too large, the cell will break apart.
In contrast, when excessive amounts of water leave a red blood cell, the cell shrinks, or crenates. This has the effect of concentrating the solutes left in the cell, making the cytosol denser and interfering with diffusion within the cell. The cell’s ability to function will be compromised and may also result in the death of the cell.
Various living things have ways of controlling the effects of osmosis—a mechanism called osmoregulation. Some organisms, such as plants, fungi, bacteria, and some protists, have cell walls that surround the plasma membrane and prevent cell lysis in a hypotonic solution. The plasma membrane can only expand to the limit of the cell wall, so the cell will not lyse. In fact, the cytoplasm in plants is always slightly hypertonic to the cellular environment, and water will always enter a cell if water is available. This inflow of water produces turgor pressure, which stiffens the cell walls of the plant. In nonwoody plants, turgor pressure supports the plant. Conversly, if the plant is not watered, the extracellular fluid will become hypertonic, causing water to leave the cell. In this condition, the cell does not shrink because the cell wall is not flexible. However, the cell membrane detaches from the wall and constricts the cytoplasm. This is called plasmolysis. Plants lose turgor pressure in this condition and wilt.
Many marine invertebrates have internal salt levels matched to their environments, making them isotonic with the water in which they live. Fish, however, must spend approximately five percent of their metabolic energy maintaining osmotic homeostasis. Freshwater fish live in an environment that is hypotonic to their cells. These fish actively take in salt through their gills and excrete diluted urine to rid themselves of excess water. Saltwater fish live in the reverse environment, which is hypertonic to their cells, and they secrete salt through their gills and excrete highly concentrated urine.
In vertebrates, the kidneys regulate the amount of water in the body. Osmoreceptors are specialized cells in the brain that monitor the concentration of solutes in the blood. If the levels of solutes increase beyond a certain range, a hormone is released that retards water loss through the kidney and dilutes the blood to safer levels. Animals also have high concentrations of albumin, which is produced by the liver, in their blood. This protein is too large to pass easily through plasma membranes and is a major factor in controlling the osmotic pressures applied to tissues.
A doctor injects a patient with what the doctor thinks is an isotonic saline solution. Do you think the solution the doctor injected was really isotonic?
The principal force driving movement in diffusion is the __________.
- particle size
- concentration gradient
- membrane surface area
What problem is faced by organisms that live in fresh water?
- Their bodies tend to take in too much water.
- They have no way of controlling their tonicity.
- Only salt water poses problems for animals that live in it.
- Their bodies tend to lose too much water to their environment.
Why does water move through a membrane?
The combined gradient that affects an ion includes its concentration gradient and its electrical gradient. A positive ion, for example, might tend to diffuse into a new area, down its concentration gradient, but if it is diffusing into an area of net positive charge, its diffusion will be hampered by its electrical gradient. When dealing with ions in aqueous solutions, a combination of the electrochemical and concentration gradients, rather than just the concentration gradient alone, must be considered. Living cells need certain substances that exist inside the cell in concentrations greater than they exist in the extracellular space. Moving substances up their electrochemical gradients requires energy from the cell. Active transport uses energy stored in ATP to fuel this transport. Active transport of small molecular-sized materials uses integral proteins in the cell membrane to move the materials: These proteins are analogous to pumps. Some pumps, which carry out primary active transport, couple directly with ATP to drive their action. In co-transport (or secondary active transport), energy from primary transport can be used to move another substance into the cell and up its concentration gradient.
Active transport mechanisms require the use of the cell’s energy, usually in the form of adenosine triphosphate (ATP). If a substance must move into the cell against its concentration gradient—that is, if the concentration of the substance inside the cell is greater than its concentration in the extracellular fluid (and vice versa)—the cell must use energy to move the substance. Some active transport mechanisms move small-molecular weight materials, such as ions, through the membrane. Other mechanisms transport much larger molecules.
Moving Against a Gradient
To move substances against a concentration or electrochemical gradient, the cell must use energy. This energy is harvested from ATP generated through the cell’s metabolism. Active transport mechanisms, collectively called pumps, work against electrochemical gradients. Small substances constantly pass through plasma membranes. Active transport maintains concentrations of ions and other substances needed by living cells in the face of these passive movements. Much of a cell’s supply of metabolic energy may be spent maintaining these processes. (Most of a red blood cell’s metabolic energy is used to maintain the imbalance between exterior and interior sodium and potassium levels required by the cell.) Because active transport mechanisms depend on a cell’s metabolism for energy, they are sensitive to many metabolic poisons that interfere with the supply of ATP.
Two mechanisms exist for the transport of small-molecular weight material and small molecules. Primary active transport moves ions across a membrane and creates a difference in charge across that membrane, which is directly dependent on ATP. Secondary active transport describes the movement of material that is due to the electrochemical gradient established by primary active transport that does not directly require ATP.
Carrier Proteins for Active Transport
An important membrane adaption for active transport is the presence of specific carrier proteins or pumps to facilitate movement: there are three types of these proteins or transporters. A uniporter carries one specific ion or molecule. A symporter carries two different ions or molecules, both in the same direction. An antiporter also carries two different ions or molecules, but in different directions. All of these transporters can also transport small, uncharged organic molecules like glucose. These three types of carrier proteins are also found in facilitated diffusion, but they do not require ATP to work in that process. Some examples of pumps for active transport are Na+-K+ ATPase, which carries sodium and potassium ions, and H+-K+ ATPase, which carries hydrogen and potassium ions. Both of these are antiporter carrier proteins. Two other carrier proteins are Ca2+ ATPase and H+ ATPase, which carry only calcium and only hydrogen ions, respectively. Both are pumps.
Figure 9: A uniporter carries one molecule or ion. A symporter carries two different molecules or ions, both in the same direction. An antiporter also carries two different molecules or ions, but in different directions. (credit: modification of work by “Lupask”/Wikimedia Commons)
Primary Active Transport
In primary active transport, the energy is derived directly from the breakdown of ATP. Often times, primary active transport such as that shown below which functions to transport sodium and potassium ions allows secondary active transport to occur (discussed in the section below). The second transport method is still considered active because it depends on the use of energy from the primary transport.
Figure 10: Primary active transport moves ions across a membrane, creating an electrochemical gradient (electrogenic transport). (credit: modification of work by Mariana Ruiz Villareal)
One of the most important pumps in animal cells is the sodium-potassium pump (Na+-K+ ATPase), which maintains the electrochemical gradient (and the correct concentrations of Na+ and K+) in living cells. The sodium-potassium pump moves K+ into the cell while moving Na+ out at the same time, at a ratio of three Na+ for every two K+ ions moved in. The Na+-K+ ATPase exists in two forms, depending on its orientation to the interior or exterior of the cell and its affinity for either sodium or potassium ions. The process consists of the following six steps.
- With the enzyme oriented towards the interior of the cell, the carrier has a high affinity for sodium ions. Three ions bind to the protein.
- ATP is hydrolyzed by the protein carrier and a low-energy phosphate group attaches to it.
- As a result, the carrier changes shape and re-orients itself towards the exterior of the membrane. The protein’s affinity for sodium decreases and the three sodium ions leave the carrier.
- The shape change increases the carrier’s affinity for potassium ions, and two such ions attach to the protein. Subsequently, the low-energy phosphate group detaches from the carrier.
- With the phosphate group removed and potassium ions attached, the carrier protein repositions itself towards the interior of the cell.
- The carrier protein, in its new configuration, has a decreased affinity for potassium, and the two ions are released into the cytoplasm. The protein now has a higher affinity for sodium ions, and the process starts again.
Several things have happened as a result of this process. At this point, there are more sodium ions outside of the cell than inside and more potassium ions inside than out. For every three ions of sodium that move out, two ions of potassium move in. This results in the interior being slightly more negative relative to the exterior. This difference in charge is important in creating the conditions necessary for the secondary process. The sodium-potassium pump is, therefore, an electrogenic pump (a pump that creates a charge imbalance), creating an electrical imbalance across the membrane and contributing to the membrane potential.
Link to Learning
Visit the site to see a simulation of active transport in a sodium-potassium ATPase.
Secondary Active Transport (Co-transport)
Secondary active transport brings sodium ions, and possibly other compounds, into the cell. As sodium ion concentrations build outside of the plasma membrane because of the action of the primary active transport process, an electrochemical gradient is created. If a channel protein exists and is open, the sodium ions will be pulled through the membrane. This movement is used to transport other substances that can attach themselves to the transport protein through the membrane. Many amino acids, as well as glucose, enter a cell this way. This secondary process is also used to store high-energy hydrogen ions in the mitochondria of plant and animal cells for the production of ATP. The potential energy that accumulates in the stored hydrogen ions is translated into kinetic energy as the ions surge through the channel protein ATP synthase, and that energy is used to convert ADP into ATP.
If the pH outside the cell decreases, would you expect the amount of amino acids transported into the cell to increase or decrease?
Injection of a potassium solution into a person’s blood is lethal; this is used in capital punishment and euthanasia. Why do you think a potassium solution injection is lethal?
If the pH outside the cell decreases, would you expect the amount of amino acids transported into the cell to increase or decrease?
Active transport must function continuously because __________.
- plasma membranes wear out
- not all membranes are amphiphilic
- facilitated transport opposes active transport
- diffusion is constantly moving solutes in opposite directions
How does the sodium-potassium pump make the interior of the cell negatively charged?
- by expelling anions
- by pulling in anions
- by expelling more cations than are taken in
- by taking in and expelling an equal number of cations
What is the combination of an electrical gradient and a concentration gradient called?
- potential gradient
- electrical potential
- concentration potential
- electrochemical gradient
Where does the cell get energy for active transport processes?
How does the sodium-potassium pump contribute to the net negative charge of the interior of the cell?
Lecture 05: Membranes and transport - Biology
C2006/F2402 '05 OUTLINE FOR LECTURE #5 Last updated 02/01/05 05:04 PM
(c) 2005 Dr. Deborah Mowshowitz , Columbia University, New York, NY
1. Kinetics and Properties of each type of Transport -- How you tell the cases apart.
A. Simple Diffusion (Case 1) Curves #1 & #2 refer to diagrams on top of handout 4A.
1. Curve #1 (uptake or concentration of substance X inside plotted vs. time) plateaus at [X]in = [X]out.
2. Curve #2 (uptake of X plotted vs concentration of X added outside) does not saturate.
3. Energy: Rxn ( X in <--> X out) is strictly reversible. (Keq = 1 standard free energy change = 0 at equil. [X]in = [X]out).
Actual free energy change and direction of transport depends on concentration of X. If [X] is higher outside, X will go in and vice versa.
4. Importance . Used by steroid hormones, some small molecules, gases. Only things that are very small or nonpolar can use this mechanism to cross membranes. Materials (usually small molecules) can diffuse into capillaries by diffusing through the liquid in the spaces between the cells. (The cells surrounding capillaries do not have tight junctions, except in the brain.)
B. Carrier mediated Transport = Facilitated Diffusion using a carrier protein (Case 3)
1. Curve #1 same as above.
2. Curve #2 saturates. See Becker fig. 8-6, or Purves 5.11 (5.10 in 5th ed. not in 6th)
3. Mechanism: Carrier acts like enzyme or permease, with Vmax, Km etc. See Becker fig. 8-8.
4. Energy as above -- substance flows down its gradient, so transport is reversible, depending on relative concentrations in and out.
5. Regulation: Activity of transport proteins can be regulated at least 3 ways. Methods a & b are common to many proteins and are only listed here for comparison (details elsewhere). Method c is unique to transmembrane proteins. (This section is about regulation of activity of pre-existing levels of protein. Synthesis and therefore protein levels are also regulated, as will be explained later.)
a. allosteric feedback inhibition/activation of carrier proteins
b. covalent modification (reversible) of the carrier proteins -- common modifications are addition and removal of phosphates see example below (Na + /K + pump)
c. removal/insertion of carrier into membranes .
(1). Newly made membrane proteins are inserted into the membrane of a vesicle, by a mechanism to be discussed later.
(2). Vesicle can fuse with plasma membrane process is reversible.
(a). Fusion of the vesicle with the plasma membrane inserts transport protein into plasma membrane where it can promote transport.
(b). Budding (endocytosis) of a vesicle back into the cytoplasm removes the transport protein and stops transport.
(3). Some channels and/or carrier proteins are regulated in this way -- channel or carrier proteins can be inserted into the membrane (or removed) in response to the appropriate hormonal signals. (An example next time.)
To see how you analyze uptake, try problem 2-1. To summarize everything so far, try 2-4.
1. Curve #1 -- Same as above except
a. Very high rate of transport -- Initial slope of Curve #1 very steep.
b. Channels often conduct ions. This has consequences. Curve #1 plateaus as above with [X]in = [X]out only if X is neutral or there is no electric potential -- see point 4 below.
2. Curve #2 : Shape like simple diffusion (linear, no saturation) at physiological concentrations. (Curve plateaus only at extraordinarily high concentrations, so we are assuming no saturation observed -- extremely high concentrations of X not reached.)
3. Mechanism. Lack of saturation and high rate of transport indicate that max. capacity of channel is very large and is not easily reached. This is explained by one or both of the following:
a. Binding of ion to channel protein is weak (Km >> 1), and/or
b. No major conformational change of channel protein is required for ion to pass through.
See Purves 44.5 (44.6) for comparison of ion pumps and ion channels Becker p. 203 (209) for comparison of carrier and channel proteins. Note that channels are very specific in spite of features a & b -- each channel transports only one or a very small # of related substances. (Mechanism of specificity has been recently figured out for one channel -- see Purves 5.10 or click on link for a nice picture of a channel and an explanation of how K + channels can be selective.)
4 . Terminology. Diffusion through a channel is usually called "facilitated diffusion" because a protein is needed (to form the channel) for transport across the membrane. However, diffusion though a channel is also sometimes called "simple diffusion," because the rate of transport as a function of [X] is generally linear, as for simple diffusion, as explained in point #2 above. In other words, the kinetics of passage through a channel are linear (at physiological concentrations of X), like simple diffusion -- not hyperbolic, as in carrier mediated transport or standard enzyme catalyzed reactions. Perhaps the best term for transport through a channel is "channel mediated diffusion."
a. Some Channels are gated = % time any particular gate is open is controlled (but each individual gate is either open all the way or shut)
(1). Ligand gated -- opens or shuts in response to ligands (= chemicals that bind to substance under discussion). Typical substances that open ligand gated channels are hormones, neurotransmitters, etc. For a picture see Purves 5.9.
(2). Voltage gated -- opens or shuts in response to changes in voltage. Allows transmission of electrical signals as in muscle and nerve -- see Becker figs. 9-9 & 9-10.
(3). Mechanically gated -- opens or shuts in response to pressure. Important in touch, hearing and balance.
b. Some channels are open all the time (ungated) An example = K + leak channels. These allow a little K + to leave or "leak out" of cells, causing cells to have a slight overall negative charge. This is critical to conduction of impulses by nerve and muscle as will be explained in detail later. Why do leak channels only allow "a little" K + to leave? See below.
7. Most channels are ion channels -- transport charged particles, not neutral molecules. This raises new energy considerations:
a. Role of charge : If X is charged, need to consider both chemical gradient & voltage (charge gradient). These can both "push" ions the same way or in opposite directions.
b. Result of charge: Keq not usually 1 here -- Curve #1 plateaus when chemical gradient and voltage are balanced (not necessarily at [X]out = [X]in). Example: K + ions stop leaking out of the cell and you reach equilibrium for K + when the charge difference across the cell membrane (which pushes K + in) balances out the concentration difference across the membrane (which pushes K + out).
See problem 2-6, A. Can you rule out transport through a channel?
D. Active Transport (Cases 4 & 5)
1. Curve #1 : when it plateaus, [X]in greater than [X]out -- because movement of substance linked to some other energy releasing reaction. (This assumes we are following the reaction Xout --> X in)
2. Curve #2 saturates. Enzyme-like protein involved -- acts as transporter or pump.
3. Energy: Not readily reversible Keq not = 1 and standard delta G not = zero. Overall reaction usually has large, negative standard delta G because in overall reaction transport of X (uphill, against the gradient) is coupled to a very downhill reaction. The downhill reaction is either
a. Splitting of ATP (in primary active transport), or
b. Running of some ion (say Y) down its gradient (in secondary active transport).
4 . Secondary (Indirect) Active Transport -- How does ATP fit in? Process occurs in 2 steps:
a. Step 1. Preparatory stage: Splitting of ATP sets up a gradient of some ion (say Y), usually a cation.
b. Step 2. Secondary Active Transport Proper: Y runs down its gradient, and the energy obtained is used to drive X up its gradient. See Becker fig. 8-10.
c. Overall: Step (1) is primary active transport step (2) is secondary and can go on (in the absence of ATP) until the Y gradient is dissipated. Note that step (1) cannot occur at all without ATP but step (2) can continue without any ATP (for a while).
5. How do you tell the two types apart? Primary is directly dependent on splitting of ATP secondary will continue even in the absence of ATP until the gradient of Y runs down.
Try problem 2-2.
6. Some Examples & Possible mechanisms (see handout 5B, texts & animations for models). Click on links for animations.
Type of Active Transport
7. Important features of Na + /K + pump (See handout)
a. Enzyme has 2 forms: one faces in (E1), one faces out (E2).
b. Forms have different affinities for K + and Na + . (See handout)
c . Role of Phosphate: addition/removal of phosphate switches the enzyme from one form to the other.
d. Role of enzymes
(1). Use of kinases & phosphatases (for addition/removal of phosphates) is a common way of regulating enzyme activity by reversible covalent modification.
(2). Enzymatic reactions:
(a). phosphorylation -- addition of phosphate groups -- catalyzed by kinases.
Kinases catalyze: X + ATP --> X-P + ADP
(b). dephosphorylation -- removal of phosphate groups -- catalyzed by phosphatases.
Phosphatases catalyze: X-P + H2O --> X + Pi
P (bold) = phosphate group Pi = inorganic phosphate (in solution)
(3). Result of enzyme activity in this case:
Kinase activity (catalyzes phosphorylation) flips enzyme "out" ( E1 --> E2)
Phosphatase activity (catalyzes removal of phosphate) flips enzyme "in" (E2 --> E1)
e. Location of Enzymes: kinase and phosphatase) are part of the pump itself. Not separate proteins.
Binding of Na + on inside activates kinase. Flips enzyme out, dumps Na + , picks up K + .
Binding of K + on outside activates phosphatase. Flips enzyme in, dumps K + , picks up Na + .
g. Stochiometry: 3 Na + out per 2 K + in. Some of charge differential balanced by Cl - transport. Cells are negative on inside relative to outside, but most of charge imbalance is NOT due to pump.
8. Important Features of Na+/Glucose co-transport
a. Enzyme has 2 forms with different affinities for glucose.
b. Role of Na+: Binding of Na + switches the protein from one form to the other alters affinity for glucose.
c. Binding of glucose probably flips protein so it faces the other way loss of glucose does the reverse.
d. Direction of glucose transport & reversibility: Either form (with or w/o Na + ) can face in or out. In normal cell, glucose always goes into the cell with Na + . Why?
9. Are pumps reversible?
a. Theoretically, all pumps (like Na + /K + pump) are reversible -- a pump can break down ATP and use the energy to drive ions up their gradient, or (if ion gradient is large enough) ions running down their gradient can provide enough delta G to drive phosphorylation of ADP to ATP. Therefore, proteins that catalyze active transport are sometimes called "ATPases" or pumps, whether their normal function is to hydrolyze ATP or to synthesize ATP.
b. Practically speaking, inside cells, most pumps are irreversible. Most (but not all) individual "pump" proteins work only one way in cells, because the standard delta G for the "usual" direction is very negative. Therefore it takes very high concentrations of products (very high ATP or very high ion concentrations, depending on the reaction) to push the reaction in the "reverse" direction. The concentrations needed to reverse the reaction are not reached in cells, but can be achieved in test tubes (by adding ATP, setting up ion gradients, etc.). So in vitro (in test tubes), but not in vivo (in living cells), you can make the pumps run in either direction. Two examples of important pumps that are reversible (in vitro), but usually run in one direction (in vivo):
(1). In the inner membranes of mitochondria and chloroplasts, chemical or light energy is used via electron transport to set up a proton gradient, which then runs down driving phosphorylation of ATP. So these systems almost always act to make ATP while ions run down their gradient. (Diff. proteins used in the two organelles.)
(2). The Na + /K + pump in the plasma membrane almost always uses up ATP -- this system drives ions up their gradients at the expense of ATP.
For more examples, see Becker table 8-3.
Now try problems 2-3 & 2-5.
II. Putting all the Methods of Transport of Small Molecules Together or What Good is All This?
A. How glucose gets from lumen of intestine --> muscle and adipose cells. An example of how the various types of transport are used. (Handout 5A and Becker fig. 11-22) Steps in the process:
1. How glucose exits lumen. Glucose crosses apical surface of epithelial cells primarily by Na + /Glucose co-transport. (2 o act. transport)
2. Role of Na+/K+ pump. Pump in basolateral (BL) surface keeps Na + in cell low, so Na + gradient favors entry of Na + . (1 o act. transport)
3. How glucose exits epithelial cells. Glucose (except that used for metabolism of epithelial cell) exits BL surface of cell (and enters interstitial fluid) by facilitated diffusion = carrier mediated transport. (Interst. fluid = fluid in between body cells.)
4. How glucose enters and leaves capillaries -- by simple diffusion through spaces between the cells. Note: this is NOT by diffusion across a membrane.
5. How glucose enters body cells -- by facilitated diffusion (= carrier mediated transport). Carrier is only "mobilized" that is, inserted into membrane (by fusion of vesicles as explained previously) in some cell types (adipose & muscle) in presence of insulin. Carrier is permanently in cell membrane in other cell types (brain, liver). See below on GLUT transporters.
6. Role of glucose phosphorylation. Conversion of G --> G-6-phosphate traps G inside cells.
For additional examples of the uses of the various types of transport processes, see Becker fig. 8-1 & 8-2.
B. How Glucose Reaches Body Cells -- Another look at handout 5-A. -- The steps in the process are described above in the order in which they occur. Here, the focus is on the various types of transport involved.
1 . Role of Active transpo rt -- Needed to get glucose from lumen to inside of epithelial cell.
a. Primary active transport -- Na + /K + pump keeps intracellular [Na + ] low.
b. Secondary active transport -- Glucose enters epithelial cells by Na + /Glucose co-transport
2. Role of Passive Transport & Phosphorylation
a. Passive Transport -- Used to move glucose the rest of the way -- out of epithelial cells, in & out of capillaries, and into body cells.
b. Phosphorylation of glucose -- Used in the body cells to keep the free glucose level at the "end of the road" low, and ensure that the glucose gradient is "downhill" from epithelial cells to capillaries to body cells.
3. Role of Diffusion: Glucose and other small molecules (but not macromolecules) diffuse in and out of capillaries through the liquid filled spaces between the cells, not by diffusing across the cell membrane. Note that proteins are too big to enter or leave capillaries this way.
4. Role of GLUT transporters (another protein/gene family)
a. GLUT proteins are responsible for carrier mediated transport of glucose. All passive glucose transport across membranes depends on a family of proteins called GLUT 1, GLUT 2, etc. This family of genes and transport proteins is responsible for all carrier mediated transport of glucose.
b. Different family members (genes and proteins) are expressed in different cell types. GLUT 1 protein is found in plasma membrane of RBC & most other cells, GLUT 2 protein on BL surface of intestinal epithelial cells, GLUT 4 protein in muscle and adipose, etc. (Note all genes for all proteins are present in all these cell types -- DNA is the same!)
c. All the genes and corresponding proteins are similar, but have significant structural and functional differences. This is another example of a gene/protein family. All the proteins have a similar overall structure -- 12 transmembrane segments, COOH and amino ends on intracellular side of membrane, etc.
d. Position & Action of GLUT 4 is insulin dependent. GLUT 4 is the only insulin dependent member of the family. Insulin triggers insertion of GLUT 4 protein into the plasma membrane, by triggering vesicle fusion, as explained above. All the other proteins are located constitutively in their respective membranes.
e. Direction of transport. Note that one member of this family (GLUT 2) is responsible for ferrying glucose OUT of epithelial cells different members are responsible for helping glucose ENTER most other cells. All family members bind glucose on one side of the membrane, change conformation and release glucose on the other side of the membrane. Which way the glucose goes (overall) depends on the relative concentrations of glucose on the two sides of the respective membrane, not on which GLUT protein is used.
Try problem 2-9.
Next time: How do large molecules cross membranes? Then how do newly made proteins get to the right place?