BIOMEMBRANE

LECTURE 4
 Membrane Transporters
 Transporters (membrane transport/carrier proteins) are specialized
membrane-spanning proteins that assist in the movement of ions,
peptides, small molecules, lipids and macromolecules across a biological
membrane.
 A carrier is not open simultaneously to both the extracellular and
intracellular environments. Either its inner gate is open, or outer gate is
open.
 In contrast, a channel can be open to both environments at the same
time, allowing the molecules to diffuse without interruption.
 Carriers have binding sites, but pores and channels do not.
 When a channel is opened, millions of ions can pass through the
membrane per second, but only 100 to 1000 molecules typically pass
through a carrier molecule in the same time.
 Each carrier protein is designed to recognize only one substance or one
group of very similar substances.
 Research has correlated defects in specific carrier proteins with specific
diseases.
Types of Ion Transporters
Mobile Ion Carriers
 Mobile ion carriers bind to a particular ion, protecting its
charge from the environment around it.
 Thus facilitating its passage into the hydrophobic interior of
the lipid membrane.
 Mobile ion carriers lose their capacity to transport ions
under conditions of very low temperatures or below the
freezing point of a membrane.
Channel Formers
 Channel formers act by introducing a hydrophilic pore into the
membrane, allowing the ion to pass, avoiding contact with the
hydrophobic interior of the membrane.
 Channel forming ionophores can maintain their ion transport
capacity even in low temperature conditions that may inhibit
mobile ion carriers.
 The average opening time of a channel is approximately one
second.
 However, if an intense gradient is added, the channel former
can transport about 20,000 cations per channel open per
millisecond.
 This value is a thousand times greater than the number of
cations that can pass in the same time interval for a single
mobile ion carrier molecule.
 Membrane Ion Channels
 Ion channels are pores that allow specific charged particles to cross the
membrane in response to an existing concentration gradient.
 Proteins are capable of spanning the cell membrane, including its hydrophobic
core, and can interact with the charge of ions because of the varied properties
of amino acids found within specific domains or regions of the protein
channel.
 Hydrophobic amino acids are found in the domains that are apposed to the
hydrocarbon tails of the phospholipids.
 Hydrophilic amino acids are exposed to the fluid environments of the
extracellular fluid and cytosol.
 Additionally, the ions will interact with the hydrophilic amino acids, which
will be selective for the charge of the ion. Channels for cations (positive ions)
will have negatively charged side chains in the pore.
 Channels for anions (negative ions) will have positively charged side chains in
the pore.
 This is called electrochemical exclusion, meaning that the channel pore is
charge-specific.
 Ion channels can also be specified by the diameter of the
pore.
 The distance between the amino acids will be specific for the
diameter of the ion when it dissociates from the water
molecules surrounding it.
 Because of the surrounding water molecules, larger pores are
not ideal for smaller ions because the water molecules will
interact, by hydrogen bonds, more readily than the amino acid
side chains.
 This is called size exclusion. Some ion channels are selective
for charge but not necessarily for size, and thus are called
a nonspecific channel.
 These nonspecific channels allow cations—particularly Na+,
K+, and Ca2+—to cross the membrane, but exclude anions.
 The sodium/potassium pump requires energy in the form of
adenosine triphosphate (ATP), so it is also referred to as an
ATPase.
 As was explained in the Membrane Transport chapter, the
concentration of Na+ is higher outside the cell than inside, and
the concentration of K+ is higher inside the cell than outside.
 That means that this pump is moving the ions against the
concentration gradients for sodium and potassium, which is
why it requires energy.
 In fact, the pump basically maintains those concentration
gradients.
 Ion channels do not always freely allow ions to diffuse across
the membrane.
 Some are opened by certain events, meaning the channels
are gated. So another way that channels can be categorized is
on the basis of how they are gated.
 Although these classes of ion channels are found in large
numbers in the cells of nervous and muscular tissue, many
also can be found in the cells of epithelial and connective
tissues.
 Ligand-Gated Channels
 A ligand-gated channel opens because a signaling molecule, a ligand,
binds to the extracellular region of the channel.
 This type of channel is also known as an ionotropic receptor because
when the ligand, known as a neurotransmitter in the nervous system, binds
to the protein, ions cross the membrane changing its charge.
 Ligand-gated channels are found in highest numbers in the dendrites and
body of the neuron.

 When the ligand, in this case the neurotransmitter acetylcholine, binds to a specific
location on the extracellular surface of the channel protein, the pore opens to allow
select ions through.
 The ions, in this case, are cations of sodium, calcium, and potassium
 Mechanically-Gated Channels
 A mechanically-gated channel opens because of a physical distortion of
the cell membrane. Many channels associated with the sense of touch
(somatosensation) are mechanically gated.
 For example, as pressure is applied to the skin, these channels open and
allow ions to enter the cell.
 Similar to this type of channel would be the channel that opens on the basis
of temperature changes, as in testing the water in the shower (Figure).

 When a mechanical change occurs in the surrounding tissue, such as pressure or touch, the
channel is physically opened.
 Thermoreceptors work on a similar principle.
 When the local tissue temperature changes, the protein reacts by physically opening the
channel.
 Voltage-Gated Channels
 A voltage-gated channel is a channel that responds to changes in the
electrical properties of the membrane in which it is embedded.
 Normally, the inner portion of the membrane is at a negative voltage.
When that voltage becomes less negative, the channel begins to
allow ions to cross the membrane (Figure).
 Some voltage-gated channels have a set voltage (threshold) above
which the channel is open and below which it is closed.
 Other types of voltage-gated channels are more complicated, opening
and closing at different voltages.
 For example, the gate may open at a specific positive voltage, but
once open will not close again until it reaches a specific negative
voltage, or vice versa.
 Making things even more complicated, some voltage-gated channels
have more than one gate where each gate responds to different
voltage properties.
 In this type of channel, both gates must be open for the ions to move
through the membrane.
 Neurons have voltage-gated channels that are specific to certain ions.
 Voltage-gated sodium and potassium channels are mostly found in the
axon hillock and axon portions of the neuron.
 Voltage-gated calcium channels are found at the axon terminal bulbs and
activate vesicle fusion with the cell membrane and neurotransmitter
release.

 Voltage-gated channels open when the transmembrane voltage changes around them.
 Amino acids in the structure of the protein are sensitive to charge and cause the pore to
open or close.
 Leakage Channels
 A leakage channel is randomly gated, meaning that it opens and closes at
random, hence the reference to leaking. There is no actual event that opens
the channel; instead, it has an intrinsic rate of switching between the open
and closed states.
 Leakage channels are found throughout the neuron and contribute to the
resting transmembrane voltage of the excitable membrane (Figure).
 Leakage channels also are found in autorhythmic cells, where the
membrane spontaneously depolarizes at a regular interval, for example
pacemaker cells in the heart.

 In certain situations, ions need to move across the membrane randomly.

 The particular electrical properties of certain cells are modified by the presence of this type
of channel.
 The membrane potential
 Today, we know that the frog's leg kicks
because neurons (nerve cells) carry information via
electrical signals.
 How do neurons in a living organism produce
electrical signals? At a basic level, neurons generate
electrical signals through brief, controlled changes in
the permeability of their cell membrane to particular
ions (such as Na+ and K+).
 Before we look in detail at how these signals are
generated, we first need to understand how
membrane permeability works in a resting neuron
(one that is not sending or receiving electrical
signals). [What is permeability?].
 We'll discuss how a neuron establishes and maintains a
stable voltage across its membrane – that is, a resting
membrane potential.
 Imagine taking two electrodes and placing one on the outside and the other
on the inside of the plasma membrane of a living cell.
 If you did this, you would measure an electrical potential difference, or
voltage, between the electrodes.
 This electrical potential difference is called the membrane potential.
 Because there is a potential difference across the cell
membrane, the membrane is said to be polarized.
 If the membrane potential becomes more positive than it is at
the resting potential, the membrane is said to be depolarized.
 If the membrane potential becomes more negative than it is at
the resting potential, the membrane is said to
be hyperpolarized.

• All of the electrical signals that neurons use to communicate are either depolarizations
or hyperpolarizations from the resting membrane potential.
Where does the resting membrane potential come from?
 The resting membrane potential is determined by the uneven distribution
of ions (charged particles) between the inside and the outside of the cell, and
by the different permeability of the membrane to different types of ions.
Types of ions found in neurons
 In neurons and their surrounding fluid, the most abundant ions are:
 Positively charged (cations): Sodium Na+ and potassium K+
 Negatively charged (anions): Chloride Cl- and organic anions
 In most neurons, K+ and organic anions (such as those found in proteins and
amino acids) are present at higher concentrations inside the cell than outside.
 In contrast, Na+ are usually present at higher concentrations outside the cell.
 This means there are stable concentration gradients across the membrane for
all of the most abundant ion types.
 How ions cross the membrane
 Because they are charged, ions can't pass directly through the hydrophobic ("water-
fearing") lipid regions of the membrane.
 Instead, they have to use specialized channel proteins that provide a hydrophilic
("water-loving") tunnel across the membrane.
 Some channels, known as leak channels, are open in resting neurons. Others are
closed in resting neurons and only open in response to a signal.
 Some ion channels are highly selective for one type of ion, but others let various
kinds of ions pass through.
 Ion channels that mainly allow K+ to pass are called potassium channels, and ion
channels that mainly allow Na+ are called sodium channels. [What about Cl- and
organic anions?]
 In neurons, the resting membrane potential depends mainly on movement
of K+ through potassium leak channels. Let's see how this works.
 What happens if only K+ can cross the membrane?
 The membrane potential of a resting neuron is primarily determined by the
movement of K+ ions across the membrane.
 So, let's get a feeling for how the membrane potential works by seeing what would
happen in a case where only K+ can cross the membrane.
 We'll start out with K+ at a higher concentration inside the cell than in the
surrounding fluid, just as for a regular neuron.
 (Other ions are also present, including anions that counterbalance the positive
charge on K+, but they will not be able to cross the membrane in our example.)
 [Where are the Na+ and Cl- ions?]
 If potassium channels in the membrane open, K+ will begin to
move down its concentration gradient and out of the cell.
 Every time a K+ ion leaves the cell, the cell's interior loses a
positive charge.
 Because of this, a slight excess of positive charge builds up on the
outside of the cell membrane, and a slight excess of negative
charge builds up on the inside.
 That is, the inside of the cell becomes negative relative to the
outside, setting up a difference in electrical potential across the
membrane.
 For ions (as for magnets), like charges repel each other and
unlike charges attract.
 So, the establishment of the electrical potential difference
across the membrane makes it harder for the remaining K+
ions to leave the cell.
 Positively charged K+ ions will be attracted to the free
negative charges on the inside of the cell membrane and
repelled by the positive charges on the outside, opposing
their movement down the concentration gradient.
 The electrical and diffusional forces that influence
movement of K+ across the membrane jointly form
its electrochemical gradient (the gradient of potential
energy that determines in which direction K+ will flow
spontaneously).
 Eventually, the electrical potential difference across the cell
membrane builds up to a high enough level that the electrical
force driving K+ back into the cell is equal to the chemical
force driving K+ out of the cell.
 When the potential difference across the cell membrane
reaches this point, there is no net movement of K+ in either
direction, and the system is considered to be in
equilibrium.
 Every time one K+ leaves the cell, another K+ will enter it.
 The equilibrium potential
 The electrical potential difference across the cell membrane that
exactly balances the concentration gradient for an ion is known as
the equilibrium potential.
 Because the system is in equilibrium, the membrane potential will
tend to stay at the equilibrium potential.
 For a cell where there is only one permeant ionic species (only one
type of ion that can cross the membrane), the resting membrane
potential will equal the equilibrium potential for that ion.
 The steeper the concentration gradient is, the larger the electrical
potential that balances it has to be.
 You can get an intuitive feeling for this by imagining the ion
concentrations on either side of the membrane as hills of different
sizes and thinking of the equilibrium potential as the force you'd need
to exert to keep a boulder from rolling down the slopes between
them.
• If you know the K+ concentration on both sides
of the cell membrane, then you can predict the
size of the potassium equilibrium potential.
Does membrane potential equal equilibrium potential?
 In glial cells, which are the support cells of the nervous
system, the resting membrane potential is equal to
the equilibrium potential.
 In neurons, however, the resting membrane potential is
close but not identical to the K+ equilibrium potential.
 Instead, under physiological conditions (conditions like
those in the body), neuron resting membrane potentials
are slightly less negative than the K+ equilibrium
potential.
 What does that mean? In a neuron, other types of ions
besides K+ must contribute significantly to the resting
membrane potential.
 Both K+ and Na+ contribute to resting potential in neurons
 As it turns out, most resting neurons are permeable to Na+ and Cl−
K+.
 Permeability to Na+ in particular, is the main reason why the
resting membrane potential is different from the potassium
equilibrium potential.
 Let’s go back to our model of a cell permeable to just one type of
ion and imagine that Na+ (rather than K+) is the only ion that can
cross the membrane.
 Na+ is usually present at a much higher concentration outside of a
cell than inside, so it will move down its concentration gradient
into the cell, making the interior of the cell positive relative to the
outside.
 Because of this, the sodium equilibrium potential—the electrical
potential difference across the cell membrane that exactly balances
the Na+ concentration gradient—will be positive.
 So, in a system where Na+ is the only permeant ion, the membrane
potential will be positive.
 In a resting neuron, both Na+ and K+ are permeant, or able to cross the membrane.
 Na+ will try to drag the membrane potential toward its (positive) equilibrium
potential.
 K+ will try to drag the membrane potential toward its (negative) equilibrium
potential.
 You can think of this as being like a tug-of-war. The real membrane potential will
be in between the equilibrium potential and the K+ equilibrium potential.
 However, it will be closer to the equilibrium potential of the ion type with higher
permeability (the one that can more readily cross the membrane).
 Opening and closing ion channels alters the
membrane potential
 In a neuron, the resting membrane potential is closer to the
potassium equilibrium potential than it is to the sodium
equilibrium potential.
 That's because the resting membrane is much more permeable
to K+ than to Na+.
 If more potassium channels were to open up—making it even
easier for K+ to cross the cell membrane—the membrane
would hyperpolarize, getting even closer to the potassium
equilibrium potential.
 If, on the other hand, additional sodium channels were to open
up—making it easier for Na+ to cross the membrane—the cell
membrane would depolarize toward the sodium equilibrium
potential.
 Changing the number of open ion channels provides a way to
control the cell’s membrane potential and a great way to
produce electrical signals.
 The Na+-K+ pump maintains Na+ and K+ gradients
 The Na+ and K+ concentration gradients across the membrane of the cell (and
thus, the resting membrane potential) are maintained by the activity of a protein
called the Na+-K+ ATPase, often referred to as the sodium-potassium pump.
 If the Na+-K+pump is shut down, the Na+ and K+ concentration gradients will
dissipate, and so will the membrane potential.
 [Why is a pump needed to maintain the concentration gradients?]
 Like the ion channels that allow Na+ and K+ to cross the cell membrane,
the Na+-K+ pump is a membrane-spanning protein.
 Unlike potassium channels and sodium channels, however, the Na+-K+ pump
doesn’t just give Na+ and K+ a way to move down their electrochemical
gradients.
 Instead, it actively transports Na+ and K+ against their electrochemical
gradients.
 The energy for this "uphill" movement comes from ATP hydrolysis (the splitting
of ATP into ADP and inorganic phosphate).
 For every molecule of ATP that's broke down, 3 Na+ ions are moved from the
inside to the outside of the cell, and 2 K+ ions are moved from the outside to the
inside.
 The sodium-potassium exchange pump

 Because 3 Na+ are exported for every 2 K+ brought into the cell,
the pump makes a small direct contribution to the resting
membrane potential (making it slightly more negative than it
would otherwise be).
 The pump's big contribution to the membrane potential, however,
is indirect:
 It maintains steady Na+ and K+ gradients, which give rise to the
membrane potential as Na+ and K+ move down their respective
concentration gradients through leak channels.