Transport of Substances Through

Cell Membrane
* The extracellular fluid
contains larger amounts of
sodium, chloride, calcium and
bicarbonate ions plus
nutrients such as glucose.
* The intracellular fluid
contains larger amounts of
potassium, magnesium,
phosphate, sulphate ions and
proteins
 Cell Membrane Consists of Lipid Bilayer
With Transport Proteins
* The lipid bilayer of the cell membrane constitutes a barrier
against movement of water molecules and water-soluble
substances.
* Lipid soluble substances can diffuse through the lipid bilayer.

* Integral proteins can function as transport proteins:

- Channel proteins: allow movement of water and selected ions.
- Carrier proteins: bind selectively with molecules or ions
conformational changes in carrier protein
movement of substances to the other side of the membrane.
 Diffusion Through Cell Membrane
I. Simple diffusion
* Simple diffusion means that kinetic movement of
molecules or ions occurs through a membrane channels or
through intermolecular spaces without any interaction
with carrier proteins in the membrane.
* The rate of diffusion is determined by amount of
substance available, velocity of kinetic motion, and
number and sizes of membrane channels.
* Diffusion of Lipid-Soluble Substances Through Lipid
Bilayer:
- Is determined by lipid solubility of the substance e.g. lipid
solubility of oxygen, nitrogen, carbon dioxide, and alcohols is
high, and all these substances can diffuse through cell
membrane.
- The rate of their diffusion is directly proportional to their lipid
solubility e.g. Oxygen can diffuse into the cell as the cell
membrane do not exist.
* Diffusion of Water and water-soluble substances Through
Protein Channels:
* Water passes rapidly through cell membrane via 1) protein
channels and 2) specialized protein pores called aquaporins.
- The rapidity of water diffusion through cell membrane is
astounding e.g. the total amount of water that diffuses in
each direction through RBC membrane/second is about 100
times as great as the volume of RBC itself.
* Water-soluble substances can pass through protein
channels in the same way as water depending on their sizes
i.e. as substances become larger, their penetration falls off
rapidly. For instance, the diameter of urea molecule is only
20% greater than that of water, yet its penetration through
the cell membrane is about 1000 times less than that of
water.
 Diffusion Through Protein Pores and Channels—
Selective Permeability and “Gating” of Channels
1. Selective permeability of protein channels
Selective permeability results from characteristics of channel,
such as its diameter, its shape, and nature of electrical charges
and chemical bonds along its inside surfaces.
* Potassium channels permit passage of potassium ions across
cell membrane about 1000 times more readily than they
permit passage of sodium ions. This selectivity cannot be
explained entirely by molecular diameters potassium ions
which are slightly larger than sodium ions ?.
- Potassium channels were found to have a tetrameric
structure consisting of four identical protein subunits
surrounding a central pore.
- At the top of the channel pore are pore
loops that form a narrow selectivity
filter which is lined with carbonyl
oxygens.
- When hydrated potassium ions enter
the selectivity filter, they interact with
the carbonyl oxygens and shed most of
their bound water molecules, permitting
dehydrated potassium ions to pass
through channel.
- The carbonyl oxygens are too far apart,
however, to enable them to interact
closely with the smaller sodium ions,
which are therefore effectively excluded
by the selectivity filter from passing
through the pore.
* Sodium channels
- Is only 0.3 to 0.5 nanometer in diameter.
- Its inner surfaces are lined with amino acids that are strongly
negatively charged.
- The negative charges pull small dehydrated sodium ions into
channels.
2. Gating of Protein Channels
Gating of Protein Channels is controlled in two ways:
A. Voltage gating:
* Electrical potential across cell membrane causes molecular
conformation of the gate which open or closed e.g. negative
charges on inside cell membrane cause outside sodium gates
to remain tightly closed; conversely, when the inside loses
its negative charge, sodium gates open and allow sodium to
pass inward.
* Potassium gates are on the intracellular ends of potassium
channels, and they open when the inside becomes positively
charged.
B. Chemical (ligand) gating:
* Binding of a chemical substance (a ligand) with the
channel protein causes conformational change in the protein
molecule that opens or closes the gate e.g. Acetylcholine
channel.
* Acetylcholine (a ligand) opens the gate of acetylcholine
channel, providing a negatively charged pore about 0.65 nm
in diameter that allows positive ions (sodium ions) smaller
than this diameter to pass through.
* Acetylcholine channel is important for transmission of
nerve signals from one nerve cell to another and from nerve
cells to muscle cells to cause muscle contraction.
 II. Facilitated Diffusion
* Facilitated diffusion means diffusion of a substance through
cell membrane with the help of a specific carrier protein.
* Facilitated diffusion differs
from simple diffusion in that as
the concentration of diffusing
substance increases, the rate of
simple diffusion continues to
increase proportionately, but in
facilitated diffusion, the rate of
diffusion approaches a maximum
called Vmax and cannot rise
greater than the Vmax level ?.
* Mechanism of facilitate diffusion: The transported molecule
enters the pore binds weakly to a binding “receptor” on
the inside the carrier protein conformational change of
carrier protein pore opens and molecule releases on
the opposite side of the membrane.
* The rate of transport of the
molecule can never be greater
than the rate at which the
carrier protein can undergo
change back and forth between
its two states limits the
rate of facilitated diffusion.
* Glucose crosses cell membrane by facilitated diffusion via a
family of membrane proteins called glucose transporters
(GLUT); at least 14 members. One of these, glucose transporter
4 (GLUT4), is activated by insulin, which can increase the rate of
facilitated diffusion of glucose as much as 10- to 20-fold in
insulin-sensitive tissues.

Factors That Affect Net Rate of Diffusion

1. Concentration Difference Across a Membrane:
The rate of net diffusion into the cell is directly proportional to
the concentration on the outside minus the concentration on
the inside:
Net diffusion ∝ (Co − Ci )
Co is concentration outside and Ci is concentration inside.
 2. Membrane Electrical Potential—The “Nernst Potential”:
* If an electrical potential is applied across the membrane, positive
charge attracts negative ions, whereas negative charge repels them.
Therefore, net diffusion occurs from left to right.
* After some time, large quantities of negative ions have moved to
the right, creating the condition that a concentration difference of
ions has developed in the direction opposite to electrical potential
difference.
* The concentration difference
now tends to move ions to the
left, while electrical difference
tends to move them to the right
* When the concentration
difference rises high enough,
the two effects balance each
other
* The electrical difference that will balance a given concentration
difference of univalent ions such as Na + can be determined from
Nernst equation:
EMF (mv) = ±61 log C1/C2
EMF is the electromotive force (voltage) between side 1 and side 2
of the membrane, C1 is the concentration on side 1, and C2 is the
concentration on side 2.
* Nernst equation is important in understanding transmission of
nerve impulses.

3. Pressure Difference Across the Membrane

Pressure difference occurs, for instance, at blood capillary
membrane in all tissues. The pressure is about 20 mm Hg
greater inside capillary than outside. The result is that increased
amounts of energy available to cause net movement of
molecules from high-pressure side toward low-pressure side.
III. Osmosis Across Selectively Permeable Membranes—“Net
Diffusion” of Water
* Assume that pure water on one side of cell membrane and sodium
chloride solution on the other side, and the membrane is selectively
permeable to water but less selective to sodium and chloride ions.
* Sodium and chloride displace water
molecules to left side of the membrane
reduce water concentration in the
right side of the membrane net
movement of water occurs from left to
right i.e. osmosis occurs from pure water
to sodium chloride solution.
* Definition: Osmosis means net
movement of water caused by a
concentration difference of water across a
membrane.
* Osmotic pressure:
- Osmotic pressure is the amount of pressure required to stop
osmosis.
- Osmotic pressure is determined by the number of particles per
unit volume of fluid, not by the mass of the particles.

* Osmolality: One osmole is 1 gram molecular weight of

osmotically active solute. Thus,
- 180 grams of glucose, which is 1 gram molecular weight of
glucose, is equal to 1 osmole of glucose because glucose does
not dissociate into ions.
- 58.5 grams of sodium chloride, which is 1 gram molecular
weight of sodium chloride, is equal to 2 osmoles because
sodium chloride dissociates into sodium and chloride ions.
- Consequently, a solution that has 1 osmole of solute dissolved
in each kilogram of water is said to have an osmolality of 1
osmole/kilogram.
- The normal osmolality of the extracellular and intracellular
fluids is about 300 milliosmoles/kilogram of water.

* Osmolarity: is the osmolar concentration expressed as

osmoles per liter of solution rather than osmoles per kilogram
of water.
- The quantitative differences between osmolarity and
osmolality are less than 1%.
- Measuring osmolarity is the usual practice in physiological
studies.
 Active Transport of Substances Through Membranes
* Definition:
Active Transport means transport of molecules or ions across
cell membrane against a concentration gradient.

* Types of active transport:

I. Primary active transport
II. Secondary active transport
- In primary active transport, energy is derived directly from
breakdown of ATP whereas in secondary active transport,
energy is derived secondarily from energy that has been stored
in the form of ionic concentration differences across a cell
membrane, created originally by primary active transport.
 I. Primary Active Transport
Sodium-Potassium pump
* Basic physical components of Na/K pump
- The carrier protein is a complex of two separate globular proteins
—a larger one called α subunit, and a smaller one called β subunit.
Although the function of β subunit is not known (might anchor
protein complex in the membrane), α subunit has three specific
features that are important for functioning of the pump:
1. It has three binding sites for
sodium ions on the inside.
2. It has two binding sites for
potassium ions on the outside.
3. The inside portion of α subunit
near the sodium binding sites has
ATPase activity Pump
* Mechanism of action of Na/K pump
Binding two potassium ions on the outside of the carrier protein
and three sodium ions bind on the inside activation of ATPase
function cleavage of ATP to ADP and liberating energy which
cause conformational change in the carrier protein extruding
three sodium ions to outside and two potassium ions to inside.

* Importance of Na/K pump

1. control cell volume: Inside the cell are negatively charged
proteins and other organic molecules (cannot escape from the cell)
that attract potassium, sodium and other positive ions osmosis
of water to cell interior cell swell and lysis Na/K pump
extrudes three sodium ions to outside for every two potassium
ions to cell inside. The membrane is less permeable to sodium ions
than it is to potassium ions sodium ions tend to stay outside
prevent cell lysis.
2. Electrogenic Nature of Na/K Pump:
Na/K pump extrudes three sodium ions to outside for every two
potassium ions to cell inside a net of one positive charge is
moved from cell interior to exterior for each pump cycle
creates positivity outside the cell and a deficit of positive
ions inside the cell i.e. negativity on the inside creates an
electrical potential across cell membrane which is a basic
requirement in nerve and muscle fibers for transmitting nerve
and muscle signals.
Primary Active Transport of Calcium Ions
* Calcium ions are normally maintained at a concentration
about 10,000 times in intracellular fluid less than that in
extracellular fluid.
* This is achieved by two primary active transport calcium
pumps:
- One in cell membrane that pumps calcium to outside cell.
- The other in intracellular organelles e.g. in ER that pump
calcium inside ER.
• Each pump has carrier protein that functions as ATPase as
that of sodium carrier protein.
Primary Active Transport of Hydrogen Ions
* It is important in parietal cells of stomach and in renal
tubules of kidneys:
1. At the secretory ends of parietal cells, the hydrogen ion
concentration is increased as much as a million-fold and then
is released into stomach along with chloride ions to form
hydrochloric acid.
2. In renal tubules, special intercalated cells found in late
distal tubules and cortical collecting ducts secrete hydrogen
ions from blood into urine against a concentration gradient of
about 900-fold.
 II. Secondary Active Transport
* There are two forms of secondary active transport: co-transport
and counter-transport.
1. Co-Transport of Glucose and Amino Acids Along with Sodium Ions:
- The transport carrier protein has
two binding sites on its exterior
side, one for sodium and one for
glucose.
- Concentration difference of
sodium (high outside and low
inside) provides diffusion energy for
transport.
- When sodium and glucose attach simultaneously to their binding
sites on carrier protein conformational change of carrier protein
and simultaneous transport of sodium and glucose to cell inside.
* Sodium co-transport of the amino acids occurs in the same
manner as for glucose, except that it uses a different set of
transport proteins. At least five amino acid transport proteins
have been identified, each of which is responsible for
transporting one subset of amino acids with specific
molecular characteristics.
* Sodium co-transport of glucose and amino acids occurs
through epithelial cells of intestinal tract and renal tubules of
kidneys.