TRANSPORT MECHANISMS

Biological Membranes
Passive Transport
Active Transport
Defects in channels causing
Inherited Disorders
Phospholipid Bilayer
Spanning proteins
Phospholipid Bilayer
Membrane Lipid Protein

Myelin Sheath 80% 20%

Plasma Membrane 50% 50%

Mitochondrial Inner
25% 75%
Membrane
Membrane Functions:
1.Selective Barrier (barrier to diffusion of hydrophilic m
a) Surround cells to hold enzymes and metabolites inside
b) surround organelles inside cells
2.Contain Enzyme Systems--energy metabolism
(oxidative phosphorylation, photosynthesis etc.)
3.Contain Transport Systems--bring food molecules
inside and maintain ion concentrations
4.Contain Specific Recognition Sites--for hormones etc.
5. Maintain ion concentrations of various substance
6. Control incoming and outgoing substances
7. Protect cell
Lipid Bilayer -- primary barrier, selective based
upon size and polarity of molecules
) Nonpolar molecules (O2, hydrocarbons, fatty
acids)-- bilayers are most permeable to small
nonpolar molecules
b) Small uncharged polar molecules (H2O, CO2) --
bilayers are somewhat permeable
c) Large polar molecules and ions -- bilayers are
relatively impermeable to large polar molecules and
to ions
d) Macromolecules (proteins, nucleic acids,
polysaccharides) -- cannot pass unless a special
mechanism exists (signal hypothesis).
Differentiated functions of the plasma
membrane of an epithelial cell.
Methods of Transport Across
Membranes
I. Passive Transport
1. Diffusion -passive transport - no energy
expended
Osmosis - Passive transport of water
across membrane
2. Facilitated Diffusion - Use of proteins to carry
polar molecules or ions across

II. Active Transport- requires energy to transport

molecules against a concentration gradient
energy is in the form of ATP
2 General Types of Transport
Passive transport
involves carriers, channels,
or direct diffusion through
a membrane.
always operates from
regions of greater
concentration to regions of
lesser concentration.
No external source of
energy is required.
Some subs also move in
response to a gradient but
do so through specific
channels formed by
proteins in the membrane
Some ions and molecules
can pass through the
membrane easily because
of conc. gradient
Examples of passive
transport include
Simple diffusion
Channel diffusion
Facilitated diffusion
3 Types of Transport Proteins
Diffusion
Powered by random movement of molecules
in a solution
Net movement is from regions of high
concentration to regions of low
concentration of solute
does not saturate as the concentration or
gradient changes
Diffusion of different substances do not
interfere with each other (no competition).
Diffusion
Net flux (amount of movement) is proportional
to the concentration difference and the
permeability of any barrier like a membrane.
Substances can cross membranes by diffusion if
they can dissolve in the oily interior of the
membrane (hydrophobic)
Diffusion can occur through tight junctions or
within bulk solutions.
Diffusion of water down its concentration
gradient is called osmosis.
Substances begin their journey dissolved in extracellular
water.
They bounce around randomly until they collide with a
lipid bilayer.
They then can become dissolved in the lipid bilayer.
They then may bounce around randomly within the
lipid bilayer until they collide with the other
(intracellular) side of the lipid bilayer.
They then can exit the lipid bilayer.
And become dissolved in intracellular water.
- Molecules move from area of [highest] to
areas of [lower], DOWN the concentration
gradient

- Molecules eventually reach equilibrium

Osmosis

The transport of water

across the membrane is
complex
Despite its polarity, it
can cross the membrane,
but this flow is limited.
Facilitated by
aquaporins, specialized
channels for water
The movement of water across cell membranes is fundamental to
life. Water constitutes roughly 70% of the mass of most living
organisms, so the orderly distribution of water is required to
maintain proper fluid balance within different anatomic
compartments. Although water is known to diffuse through lipid
bilayers, diffusion is not sufficiently rapid for many physiological
processes. To accommodate these needs, a family of membrane
channel proteins evolved for rapid transport of water across
biological membranes. These proteins, termed aquaporins, are
found in all life forms, including archaea, eubacteria, fungi, plants,
and all phyla of animals.
Osmosis: The human perspective
More than 11 aquaporins The human genetic
found in mammals disease, hereditary
2 classes: specific for (nephrogenic) diabetes
water only; those that insipidus (NDI) caused
allow other small by a non-functional
hydrophilic mol aquaporin protein,
(glycerol, urea) to cross causes the excretion of
the membrane large volumes of dilute
urine.
Two chambers containing different solute concentrations are separated by
a semipermeable membrane (a). In (b) water diffuses across the membrane until
the difference in height, h, equals the osmotic pressure. In (c) a piston pushes on
the liquid in the righthand chamber; the amount of pressure required to keep water
from diffusing into this chamber is equal to the osmotic pressure
Plant and Animal Cells put into
various solutions
Tonicity is a relative term
Hypotonic Solution - One solution has a lower
concentration of solute than another.
Hypertonic Solution - one solution has a
higher concentration of solute than another.
Isotonic Solution - both solutions have same
concentrations of solute.
Diffusion
Factors that affect diffusion
Temperature
-Temperature is an increase in molecular speed
(kinetic energy)
-Therefore, will higher temperature increase or
decrease the rate of diffusion?

Molecular Weight
-Larger molecules will have more drag across the
membrane thus producing a slower rate of diffusion
-Test using HCl (36.461) and NH4OH (35.050)
Facilitated Diffusion
Proteins act as carriers or pores that
permit flux of substances that cannot
diffuse directly through the membrane.
Movement is still passive (like
diffusion), from high concentration to
low.
Occurs across cell membranes only.
Saturates when substance reaches
high concentrations due to lack of
available protein (exhibit saturation)
Related substances can compete for
the same carrier or pore (Specificity)
Maximum rate of transport (fully
saturated) is called Tm, the transport
maximum.
Facilitated Diffusion
Facilitated Diffusion
Allows diffusion of large, membrane insoluble compounds such as
sugars (glucose) and amino acids
Does not require energy (passive transport)
Substance binds to membrane transport protein
Molecules may enter the cell and leave the cell through the transport
protein.
Particles move from areas of high concentration to areas of low
concentration; diffuse
Uniport (facilitated diffusion)
carriers mediate transport of a
single solute. An example is the
GLUT1 glucose carrier. The
ionophore valinomycin is also a
uniport carrier.
Calcium enters cells through 2 kinds of channels:
voltage-gated calcium channels activated by depolarization, and ligand-gated
Ca2+ Channels which in various types of cells are activated by many different
types of hormones and neurotransmitters.

Ca2+ is pumped out of the cells

in exchange for 2 H+ by a Ca2+-H+ ATPase.

It is also transported out of the cells by an antiport driven by the sodium gradient
that exchanges 3 Na+ for each Ca2+.

ICF Ca2+ : 100 nm/l

ECF Ca2+ : 1.2 nmol/l
There is a marked inwardly directed
concentration gradient and inwardly directed
electrochemical gradient.
Ligand-gated channels
Active Transport

driven by energy other than stored in the concentration

gradient or the electrochemical gradient of the transported
molecule.
requires usually the expenditure of ATP and the help of
specific transport proteins.
In this way can even large molecules can be channelled
through the membrane.
Active transport can only occur at intact, closed membranes.
 Primary Active Transport
ions are moved across a cell membrane by
carrier proteins that directly couple transport
with hydrolysis of ATP.

Such carrier proteins are usually called ATPases,

molecules that collect the free energy of ATP
hydrolysis to move ions up an electrochemical
gradient.

The only substances transported by carriers that

directly hydrolyze ATP are positively-charged ions
- Na+, K+, Ca++ or H+.
Primary Active Transport
Proteins in the membrane can also act as
pumps.
Move ions or small molecules from low
concentration to high concentration (i.e. up
their gradients).
Require cellular energy, usually as ATP
Saturates when substance reaches high
concentrations due to lack of available protein.
Example: Na-K ATPase
Present in nearly every cell in the body
Pumps 3 Na ions out in exchange for 2 K ions
pumped in (cost=1 ATP)
electrogenic, contributes directly to the
separation of charges across the membrane
Other pumps include the Ca-ATPase, and the
H-ATPase.
P-type ion pump (P- phosphorylation)
During the pumping cycle, the hydrolysis of ATP leads
to the transfer of the released PO4 group to an aspartic
acid residue of the transport protein , which in turn
causes conformational changes within the protein
 Important examples of primary
active transporters include:
Sodium pumps or Na+-K+ ATPase are undoubtedly the premier
members of this class of transport proteins.
pump 3 sodium ions out of the cell in concert with pumping 2
potassium ions into the cell, maintaining an intracellular
environment that has low sodium and high potassium (the opposite
of extracellular fluid).
In most cells, one-third to one-half of the total energy
expended is used to run these ion pumps.

In neurons, which repeatedly gain sodium during action potentials,

it's more like two-thirds of the ATP usage goes toward fueling this
single transporter. Cells die rather rapidly when ATP synthesis is
inhibited, and one of the major causes of this effect is the altered
intracellular environmment that results when Na+-K+ pumps cannot
function.
Na+/K+ -ATPase pump
Sodium ions bind to
cytoplasmic side of
protein to change its
nformation
In its new conformation, the protein
binds a molecule of ATP and cleaves
it into ADP and PO4. ADP is
released, but the PO$ is covelantly
linked to the protein. The protein is
now phosphorylated
The phosphorylation of the protein induces a scond conformational
change in the protein. This change tanslocates the 3 sodium ions
across the membrane, so they now face the exterior. In this
conformation the protein has low affinity for sodium, sodium ions
dissociates and diffuse to the ECF.
The new conformation has a high affinity
for K+, 2 of which bind to the
extracellular side of the protein as soon as
it is free of Na+ ions
The binding of K+ ions causes another
conformational change in the protein, this
time resulting in the hydrolysis of the bound
PO4 group
Freed of the PO4 group the protein reverts to its original
conformation, exposing the 2 K+ ions to the cytoplasm. This
conformation has a low affinity for for K+ so the 2 bound K+
dissociate from the protein and diffuse into the interior of the cell.
The original conformation has a low affinity for Na+; when these
ions bind, they initiate another cycle.
These pumps transport H+ only.
- found in lysosomes, endosomes and plant vacuoles.
- transport H+ ions to make the lumen or inside of the
lysosome acidic (pH 4.5 - 5.0)
- many of these pumps are paired with Cl- channels to
offset the electrical gradient that is produced by pumping
H+ across the membrane.
Proton pumps function to transport H+ ions out of the
cytoplasm of cells.

The stomach is famous for secreting acid, and the relevant

mechanism is based on a H+-K+ ATPase in the lumenal
membrane of gastric parietal cells.
This ion pump allows the parietal cell to secrete H+ ions against a
roughly million-fold concentration gradient - truly a case of active
transport!

Another example of a proton pump is found in the membranes

of lysosomes. These intracellular garbage disposals maintain a pH
of 4.5-5.0 by pumping H+ ions from the cytoplasm into the interior
of the lysosome through a carrier protein that is an ATPase.
 H+/k+-ATPase
The epithelial lining of the stomach
also contains a P-type pump, the
H+/K+-ATPase which secretes conc.
Acid, up to .16 N HCl into the stomach

The H+/K+-ATPase molecules are

present in the walls of cytoplasmic
vesicles.
Food in stomach -histamine to parietal cells-
Vesicles move to apical surface, fuse with
plasma m- acid secretion

Prilosec prevents heartburn by inhibiting the pump.

Zantac,Pepcid,Tagamet dont inhibit the pump dirrectly, but block the
receptor
On the surface of parietal cells preventing the cells from hormone
activation.
Calcium pumps

Calcium pumps are one mechanism cells use to shuttle

calcium across cell membranes. Calcium ATPases in the
plasma membrane mediate active transport of calcium
out of cells, serving to maintain the normal, very low
levels of free cytoplasmic calcium.
Another well studied example of a calcium ATPase is
found in the sarcoplasmic reticulum of muscle cells,
where it serves to pump calcium from the cytoplasm
into those specialized forms of endoplasmic reticulum
after a period of muscle contraction. Structurally, this
active transporter is very similar to the Na+-K+ ATPase.
Transporters are of two general
classes: carriers and channels.
These are exemplified by two
ionophores (ion carriers
produced by microorganisms):
Classes of Carrier Proteins
Secondary Active Transport
Secondary active transpoSecondary active transport
Secondary active transport
In secondary active transport or co-transport, energy is used to transport molecules across a
membrane; however, in contrast to primary active transport, there is no direct coupling of
ATP; instead, the electrochemical potential difference created by pumping ions out of the
cell is used.[5]
The two main forms of this are antiport and symport.
rtS
Cotransport (Secondary Active Transport)

Transport across membranes can be fueled not

only by ATP, but by the energy stored in ion
gradients.
In such cases, the free energy released during the
transport of ions down an electrochemical gradient is
used to pump other ions or molecules up their
electrochemical gradient. This process is called
cotransport because one carrier protein mediates the
transport of both species. Some cotransporters carry
both solutes in the same direction (symport), while
others transport one solute into the cell and the other
out of the cell (antiport).
SYMPORT
 Symport (cotransport) carriers
bind two dissimilar solutes
(substrates) and transport them
together across a
membrane. Transport of the two
solutes is obligatorily coupled. A
gradient of one substrate, usually
an ion, may drive uphill (against
the gradient) transport of a co-
substrate. It is sometimes referred
to as secondary active transport
Examples include the glucose-Na+ symport found in
plasma membranes of some epithelial cells the bacterial
lactose permease, a H+ symport carrier.
SYMPORT
cotransport of glucose and sodium into the small
intestinal absorptive epithelial cell. The carrier protein
that cotransports is the sodium-dependent hexose
transporter SGLUT-1. It is distributed in the apical (lumen-
facing) membrane of the cells. The basolateral membranes
of these cells contain Na+-K+ pumps, which actively export
sodium from the cell - this maintains a low intracellular
concentration of sodium and also establishes a strong
electrochemical gradient of sodium across the apical
membrane. This gradient of sodium provides the energy
for running SGLUT-1. Cotransport is achieved through a
series of conformational changes:
a sodium ion in the intestinal lumen binds to
SGLUT-1, which induces a conformational
change that allows a molecule of glucose to bind;

glucose binds, resulting in the carrier protein

reorienting in the membrane such that the
bound sodium ion and glucose face the
cytoplasm ;

glucose, then sodium dissociate from the

carrier into the cytoplasm ;

the carrier reorients back to its original

position.
ANTIPORT
ANTIPORT
Sodium-proton cotransport involves transport of a sodium
ion into the cell coupled to transport of a hydrogen ion out of
the cell; both ions are transported by the same carrier protein.
This process is particularly important in proximal tubules of
the kidney.

Sodium-calcium cotransport occurs in most if not all cell

membranes. Sodium is carried into the cell in exchange for
calcium, which is expelled from the cell.
Inc in blood glucose level insulin secretion- transl
From cytoplasm to cell surface
Stimulate glucose uptake into various target cells
(skeletal muscle/fat cells)
Insulin responsive cells share a common
Isoform of facultative glucose transporter, GLUT4

When insulin levels are low-

Lactose permease catalyzes uptake of the disaccharide
lactose into E. coli bacteria, along with H+, driven by a
proton electrochemical gradient. It is the first carrier
protein for which an atomic resolution structure has been
determined. Lactose permease has been crystallized
with thiodigalactoside (TDG), an analog of lactose.

The substrate binding site is at the apex of an aqueous

cavity between 2 domains, each consisting of 6
transmembrane a-helices. In the conformation observed in
this crystal structure, the substrate analog is accessible
only to what would be the cytosolic side of the intact
membrane.
Residues identified as being essential for H+-binding are
also near the middle of the membrane.
Antiport (exchange diffusion)
exchange one solute for another
across a membrane
exhibit "ping pong" kinetics. A
substrate binds and is ransported
across the membrane. Then nother
substrate binds and is transported
in the other direction
Only exchange is catalyzed, not
net transport
An example is the adenine
nucleotide translocase
(ADP/ATP exchanger),
 Antiport
An example is the adenine nucleotide translocase
(ADP/ATP exchanger), which catalyzes 1:1 exchange of
ADP for ATP across the inner mitochondrial membrane.