TRANSPORT ACROSS CELL MEMBRANES

LEARNING OBJECTIVES

At the end of this lecture, students should be able to:

 Define and list the various types of passive transport

 Know the difference between channel and carrier proteins and how they influence diffusion
 Understand the concept of selective permeability and the regulation of gated channels
 List and explain the factors that determine the magnitude of solute flux in a carrier-mediated
transport.
 List and explain the factors that affect the rate of diffusion
 Differentiate between the terms osmole, osmolarity, osmolality and tonicity. List the typical
value and normal range for plasma osmolarity.
 Define and list the various types of active transport, differentiate between primary and
secondary active transport with typical examples.
 Explain the mechanism of action of the Na+/K+-ATPase transporter.
 Explain endocytosis and exocytosis giving typical examples.

INTRODUCTION

The movement of water and solutes—such as ions, sugars, and other molecules—between the
extracellular and intracellular fluid is critical for the survival of all cells, tissues, and organs.
In this way, important biological molecules are delivered to cells and wastes are removed and
eliminated from the body. In addition, regulation of ion movements creates the electrical
properties that are crucial to the function of many cell types. The cells achieve these by
means of transport mechanisms across the cell membrane. Structure of the cell membrane is
well suited for the transport of substances in and out of the cell. Lipids and proteins of cell
membrane play an important role in the transport of various substances between extracellular
fluid (ECF) and intracellular fluid (ICF). Hence, it is a general principle of physiology that
controlled exchange of materials occurs between compartments and across cellular
membranes.

BASIC MECHANISMS OF CELLULAR TRANSPORT

Broadly speaking, there are two types of basic mechanisms involved in the transport of
substances across the cell membrane based on the source of energy expenditure:

1. Passive Transport
a. Simple diffusion (through lipid and protein layers)
b. Facilitated (carrier-mediated) diffusion
 Special types of passive transport
i. Bulk flow
ii. Filtration
iii. Osmosis
2. Active Transport (carrier-mediated)
a. Primary active transport
 b. Secondary active transport
 Special types of active transport
i. Endocytosis (pinocytosis and phagocytosis)
ii. Exocytosis
 Epithelial transport (paracellular and transcellular pathways)

Passive Transport
Passive transport is the transport of substances along the concentration gradient or electrical
gradient or both (electrochemical gradient). It is also known as diffusion or ‘downhill’
movement. It is called “passive” because it does not directly or indirectly use ATP, the
energy currency of cells, for the movement of substances across the cell membrane.
However, all motion or movement of substances depend on some energy or the other as we
shall see shortly.

Diffusion: One of the fundamental physical features of molecules of any substance,

whether solid, liquid, or gas, is that they are in a continuous state of movement or vibration.
The energy for this movement comes from what Physicists call “heat”; the warmer a
substance is, the faster its molecules move. The random thermal motion of molecules in a
liquid or gas will eventually distribute them uniformly throughout a container. This
movement of molecules from one location to another solely as a result of their random
thermal motion is known as simple diffusion.

a. Simple Diffusion: Means that kinetic movement of molecules or ions occurs through
a membrane opening or through intermolecular spaces without any interaction with
carrier proteins in the membrane. Simple diffusion can occur through the cell
membrane by two pathways: (1) through the interstices of the lipid bilayer if the
diffusing substance is lipid soluble and (2) through watery channels that penetrate all
the way through some of the large transport proteins.
 Diffusion of Lipid-Soluble Substances through the Lipid Bilayer
An important factor that determines how rapidly a substance diffuses through the lipid
bilayer is the lipid solubility of the substance. Oxygen, nitrogen, carbon dioxide,
alcohols, fatty acids, and steroid hormones are examples of nonpolar (lipid soluble)
molecules that diffuse rapidly through the lipid portions of membranes.
 Diffusion of Water and Other Lipid-Insoluble Molecules through Protein (Ion)
Channels
Ions such as Na+, K+, Cl-, and Ca2+ diffuse across plasma membranes at much faster
rates than would be predicted from their very low solubility in membrane lipids. Also,
different cells have quite different permeabilities to these ions, whereas nonpolar
substances have similar permeabilities in nearly all cells. Moreover, artificial lipid
bilayers containing no protein are practically impermeable to these ions; this indicates
that the protein component of the membrane is responsible for these permeability
differences. Some of these proteins form ion channels that allow ions to diffuse
across the membrane. The diameters of ion channels are very small, only slightly
 larger than those of the ions that pass through them. The small size of the channels
prevents larger molecules from entering or leaving.

A characteristic feature of the protein channels is the selective permeability. That is,
each channel can permit only one type of ion to pass through it. Accordingly, the
channels are named after the ions which diffuse through these channels such as sodium
channels, potassium channels, etc. Another characteristic feature of the protein channels
is that they are regulated by the presence of gates, either electrically (voltage-gated
channels), chemically (ligand-gated channels), or mechanically (mechanical-gated
channels)

b. Facilitated Diffusion: Like simple diffusion, facilitated diffusion occurs down an

electrochemical potential gradient; thus it requires no input of metabolic energy.
Unlike simple diffusion, however, facilitated diffusion uses a membrane carrier.
Among the most important facilitated-diffusion systems in the body are those that
mediate the transport of glucose across plasma membranes. Without such glucose
transporters, or GLUTs as they are abbreviated, cells would be virtually impermeable
to glucose, which is a polar molecule. Four factors determine the magnitude of solute
flux through a carrier mediated-transport system: (1) the solute concentration, (2) the
affinity of the transporters for the solute, (3) the number of transporters in the
membrane, (4) the rate at which the conformational change in the transport protein
occurs.

Factors That Affect the Rate of Diffusion across Membranes

I. Permeability of the Cell Membrane: Rate of diffusion is directly proportional to the
permeability of cell membrane. Since the cell membrane is selectively permeable,
only limited number of substances can diffuse through the membrane.
II. Temperature: Rate of diffusion is directly proportional to the body temperature.
Increase in temperature increases the rate of diffusion.
III. Electrochemical Gradient across the Cell Membrane: Rate of diffusion is directly
proportional to the concentration gradient or electrical gradient of the diffusing
substances across the cell membrane. However facilitated diffusion has some
limitation beyond certain level of concentration gradient.
IV. Solubility of the Substance: Diffusion rate is directly proportional to the solubility of
substances, particularly the lipid-soluble substances. Since oxygen is highly soluble in
lipids, it diffuses very rapidly through the lipid layer.
V. Thickness of the Cell Membrane: Rate of diffusion is inversely proportional to the
thickness of the cell membrane. If the cell membrane is thick, diffusion of the
substances is very slow.
VI. Size of the Molecules: Rate of diffusion is inversely proportional to the size of the
molecules. Thus, the substances with smaller molecules diffuse rapidly than the
substances with larger molecules.

 Special Types Of Passive Transport

 i. Bulk Flow: Bulk flow is the diffusion of large quantity of substances from a
region of high pressure to the region of low pressure. It is due to the pressure
gradient of the substance across the cell membrane. A good example is the
exchange of gases across the respiratory membrane in lungs.
ii. Filtration: The capillary wall separating plasma from interstitial fluid is
different from the cell membranes separating interstitial fluid from
intracellular fluid because the pressure difference across it makes filtration a
significant factor in producing movement of water and solute. By definition,
filtration is the process by which fluid is forced through a membrane or other
barrier because of a difference in pressure on the two sides.
iii. Osmosis: Osmosis is the net diffusion of water across a semipermeable
membrane because of differences in solute concentration. Concentration
differences of impermeant solutes establish osmotic pressure differences, and
this osmotic pressure difference causes water to flow by osmosis. The amount
of pressure required to stop osmosis is called the osmotic pressure. The
addition of a solute to water decreases the concentration of water in the
solution compared to the concentration of pure water. For example, if a solute
such as glucose is dissolved in water, the concentration of water in the
resulting solution is less than that of pure water. A given volume of a glucose
solution contains fewer water molecules than an equal volume of pure water
because each glucose molecule occupies space formerly occupied by a water
molecule.

The degree to which the water concentration is decreased by the addition of solute
depends upon the number of particles (molecules or ions) of solute in solution (the
solute concentration) and not upon the chemical nature of the solute.

Active Transport
Active transport uses energy to move a substance uphill across a membrane—that is,
against the substance’s concentration gradient. As with facilitated diffusion, active transport
requires a substance to bind to the transporter (carrier protein) in the membrane. Because these
transporters move the substance uphill, they are often referred to as pumps. As with facilitated-
diffusion transporters, active-transport transporters exhibit specificity and saturation—that is,
the flux via the transporter is maximal when all transporter binding sites are occupied. The net
movement of solute from lower to higher concentration and the maintenance of a higher steady-
state concentration on one side of a membrane can be achieved only with continuous input of
energy into the active-transport process. Two means of coupling energy to transporters are
known: (1) the direct use of ATP in primary active transport and (2) the use of an
electrochemical gradient across a membrane to drive the process in secondary active transport.

Carrier proteins involved in active transport are of two types:

1. Uniport: Carrier protein that carries only one substance in a single direction. It is also
known as Uniport pump.
 2. Symport (cotransport) or antiport (countertransport): Symport or antiport is the carrier
protein that transports two substances at a time. Carrier protein that transports two
different substances in the same direction is called Symport or symport pump.
Carrier protein that transports two different substances in opposite directions is called
antiport or antiport pump.

Primary Active Transport: The hydrolysis of ATP by a transporter provides the energy
for primary active transport. The transporter itself is an enzyme called ATPase that catalyzes
the breakdown of ATP and, in the process, phosphorylates itself. Phosphorylation of the
transporter protein is a type of covalent modulation that changes the conformation of the
transporter and the affinity of the transporter’s solute binding site.
One of the best-studied examples of primary active transport is the movement of sodium
and potassium ions across plasma membranes by the Na+/K+-ATPase pump. This
transporter, which is present in all cells, moves Na + from intracellular to extracellular fluid,
and K+ in the opposite direction. In both cases, the movements of the ions are against their
respective concentration gradients.

Mechanism of action of Na+-K+ pump: Three sodium ions from the cell get attached to the
receptor sites of sodium ions on the inner surface of the carrier protein. Two potassium ions
outside the cell bind to the receptor sites of potassium ions located on the outer surface of the
carrier protein. Binding of sodium and potassium ions to carrier protein activates the enzyme
ATPase. ATPase causes breakdown of ATP into adenosine diphosphate (ADP) with the
release of one high energy phosphate. Now, the energy liberated causes some sort of
conformational change in the molecule of the carrier protein. Because of this, the outer
surface of the molecule (with potassium ions) now faces the inner side of the cell. And, the
inner surface of the protein molecule (with sodium ions) faces the outer side of the cell. Now,
dissociation and release of the ions take place so that the sodium ions are released outside the
cell (ECF) and the potassium ions are released inside the cell (ICF). Exact mechanisms
involved in the dissociation and release of ions are not yet known.

In addition to the Na+/K+-ATPase transporter, the major primary active-transport

proteins found in most cells are (1) Ca2+- ATPase; (2) H+-ATPase; and (3) H+/K+-ATPase.
Together, the activities of these and other active-transport systems account for a significant
share of the total energy usage of the human body.

Secondary Active Transport: In secondary active transport, the movement of an ion

down its electrochemical gradient is coupled to the transport of another molecule, often an
organic nutrient like glucose or an amino acid. Thus, transporters that mediate secondary
active transport have two binding sites, one for an ion—typically but not always Na +—and
another for a second molecule.

The creation of a Na+ concentration gradient across the plasma membrane by the
primary active transport of Na+ is a means of indirectly “storing” energy that can then be used
to drive secondary active-transport pumps linked to Na+. Ultimately, however, the energy for
secondary active transport is derived from metabolism in the form of the ATP that is used by
the Na+/K+-ATPase to create the Na+ concentration gradient. As noted earlier, the net
movement of Na+ by a secondary active-transport protein is always from high extracellular
concentration into the cell, where the concentration of Na + is lower. Therefore, in secondary
active transport, the movement of Na+ is always downhill, whereas the net movement of the
actively transported solute on the same transport protein is uphill, moving from lower to
higher concentration. The movement of the actively transported solute can be either into the
cell (in the same direction as Na+), in which case it is known as cotransport, or out of the
cell (opposite the direction of Na+ movement), which is called countertransport. Substances
carried by sodium cotransport are glucose, amino acids, chloride, iodine, iron and urate.

 Special Types Of Active Transport

i. Endocytosis: Endocytosis is defined as a transport mechanism by which the
macromolecules enter the cell. Macromolecules (substances with larger molecules)
cannot pass through the cell membrane either by active or by passive transport
mechanism. Such substances are transported into the cell by endocytosis. Three
common types of endocytosis may occur in a cell. These are pinocytosis (“cell
drinking”), phagocytosis (“cell eating”), and receptor-mediated endocytosis.
a. Pinocytosis: In pinocytosis, also known as fluid endocytosis, an endocytotic
vesicle encloses a small volume of extracellular fluid. This process is
nonspecific because the vesicle simply engulfs the water in the extracellular
fluid along with whatever solutes are present. These solutes may include ions,
nutrients, or any other small extracellular molecule. Large macromolecules,
other cells, and cell debris do not normally enter a cell via this process
b. Phagocytosis: In phagocytosis, cells engulf bacteria or large particles such as
cell debris from damaged tissues. In this form of endocytosis, extensions of
the plasma membrane called pseudopodia fold around the surface of the
particle, engulfing it entirely. The pseudopodia, with their engulfed contents,
then fuse into large vesicles called phagosomes that are internalized into the
cell. Phagosomes migrate to and fuse with lysosomes in the cytoplasm, and the
contents of the phagosomes are then destroyed by lysosomal enzymes other
molecules. Whereas most cells undergo pinocytosis, only a few special types
of cells, such as those of the immune system, carry out phagocytosis.
a. Receptor-mediated Endocytosis: Receptor-mediated endocytosis is the
transport of macromolecules with the help of a receptor protein. Surface of
cell membrane has some pits which contain a receptor protein called clathrin.
Together with a receptor protein (clathrin), each pit is called receptor-coated
pit. These receptor-coated pits are involved in the receptor-mediated
endocytosis
ii. Exocytosis: Exocytosis is the process by which the substances are expelled from the
cell. In this process, the substances are extruded from cell without passing through the
cell membrane. This is the reverse of endocytosis. Exocytosis performs two functions
for cells: (1) It provides a way to replace portions of the plasma membrane that
endocytosis has removed and, in the process, a way to add new membrane
components as well; and (2) it provides a route by which membrane impermeable
 molecules (such as protein hormones) that the cell synthesizes can be secreted into the
extracellular fluid.

Exocytosis is the mechanism by which most neurons communicate with each other through
the release of neurotransmitters stored in secretory vesicles that merge with the plasma
membrane.

 Epithelial Transport
Epithelial cells line hollow organs or tubes and regulate the absorption or secretion of
substances across these surfaces. One surface of an epithelial cell generally faces a hollow or
fluid-filled tube or chamber, and the plasma membrane on this side is referred to as the apical
membrane (also known as the luminal membrane). The plasma membrane on the opposite
surface, which is usually adjacent to a network of blood vessels, is referred to as the
basolateral membrane (also known as the serosal membrane). The two pathways by which a
substance can cross a layer of epithelial cells are (1) the paracellular pathway, in which
diffusion occurs between the adjacent cells of the epithelium; and (2) the transcellular
pathway, in which a substance moves into an epithelial cell across either the apical or
basolateral membrane, diffuses through the cytosol, and exits across the opposite membrane.
Diffusion through the paracellular pathway is limited by the presence of tight junctions
between adjacent cells, because these junctions form a seal around the apical end of the
epithelial cells.