Membrane Transport

©Anna Jankowska
[email protected]

Department of Cell Biology

www.katbiolkom.ump.edu.pl
Cell membrane
 Role of the cell membrane
• Isolates the cytoplasm from the external environment
• Regulates the exchange of substances
• Communicates with other cells
• Controles interaction between cells
• Maintains an electrical potential difference between
cytoplasm and extracllular sites

Essential Biological Functions

• Cell metabolism
• Cell growth and differentiation
• Cell adherence
• Neurotransmission
• Immune response
 Membrane components
All membranes are made up of:
• lipids,
• proteins,
• carbohydrates.
Most plasma membranes consist of ~50% lipid and 50% protein by weight,
with the carbohydrate portions of glycolipids and glycoproteins constituting 5 to
10% of the membrane mass.
The ratio varying however from ~80% lipid (in the myelin membrane) to less than 30% lipid
(in the mitochondrial membrane).
The glycolipids are found exclusively in the outer leaflet of the plasma
membrane, with their carbohydrate portions exposed on the cell surface.

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 Phospholipid bilayer

The fundamental structure of the membrane is the

phospholipid bilayer,
which forms a stable barrier between two aqueous compartments.

Hydrogen bonds form between the phospholipid polar "heads" and the
watery environment inside and outside of the cell.
Hydrophobic interactions force the "tails" to face inward.

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Phospholipids are not bonded to each other,
which makes the double layer fluid

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Control of Membrane Fluidity
The length of the fatty acid;
The degree of unsaturation of their side chains;
The amount of cholesterol;
The temperature.

Membrane fluidity is crucial to: exoctosis, endocytosis,

membrane trafficking, and membrane biogenesis.
 Asymmetry of lipid bilayer
A common feature of eukaryotic membranes is the non-random distribution of
different lipid species in the lipid bilayer.
Lipid asymmetry provides the two sides of the plasma membrane with
different biophysical properties and influences numerous cellular functions.
The outer leaflet of the plasma membrane consists mainly of phosphatidylcholine and sphingomyelin.
Phosphatidyethanolamine, phosphatidylserine and phosphatidylinositol are the predominant
phospholipids of the inner leaflet.

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 Glycocalyx (cell coat)

A glycoprotein-polysaccharide covering that surrounds

the cell membranes of cells.
Composition: polar oligosaccharide side chains linked
covalently to most proteins and some lipids of the
plasmolemma.
It also contains proteoglicans (glycosaminoglycans bound to
integral proteins).
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The glycocalyx contributes to cell-cell recognition,
communication, and intracellular adhesion.

Functions:
• protects cells from injury by preventing contact with inappropriate substances;
• aids in attachment of some cells (fibroblasts, but nor epithelial cells) to EXM components;
• binds antigens and enzymes on the cell surface;
• facilitates cell-cell recognition;
• assists T cells and antigen-presenting cells in aligning with each other in proper fashion;
• allows to distinguish between its own healthy cells and transplanted tissues, diseased cells, or
invading organisms;
• in blood vessels, it lines the endothelial surface to decrease frictional forces as the blood rushes
by and diminishes loss of fluid from the vessel.
 Membrane proteins perform a variety of functions
vital to the survival of organisms

1. Channel Proteins – form small openings for molecules to diffuse through.

2. Carrier Proteins – binding site on protein surface "grabs" certain molecules
and pulls them into the cell.
3. Receptor Proteins – molecular triggers that set off cell responses (such as
release of hormones or opening of channel proteins).
4. Cell Recognition Proteins – ID tags, to identify cells to the body's immune
system.
5. Enzymatic Proteins – carry out metabolic reactions.
Diversity of Molecules Makes Fluid Mozaic

The fluid mosaic model of the structure of cell membranes encouraged

the study of membranes and the role of each of their components.
Membranes are formed by a fluid lipid bilayer which confers exceptional
physical properties to the cell and whose lipids interact with proteins.
 Lipid Rafts Model

Sphingolipids and cholesterol form “rafts”

that move laterally within the plasma
membrane and may associate with specific
membrane proteins.

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 Lipid rafts
• The interactions of lipids with proteins allow membranes to create a variety of
domains based on the type of lipid components.
• Those domains conform different structures and exert specific functions, such
as the propagation of different cell signals, and the uptake of extracellular
molecules by endocytosis.
• The clusters of sphingolipids and cholesterol form “rafts” that move
laterally within the plasma membrane and may associate with specific
membrane proteins.

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The raft concept of membrane subcompartmentalization.

 Lipid rafts
Functional microdomains of plasma membrane

The membrane is able to laterally segregate its constituents to

coordinate biological functions.
This capability underlies the raft concept of membrane
subcompartmentalization.
Lipid rafts are fluctuating nanoscale (20-100 nm) assemblies of
sphingolipids, cholesterol, and specific proteins that can be stabilized to
unite, forming functional platforms that regulate membrane signaling
and trafficking, or endocytosis.

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 Lipid rafts have been shown to associate with
cytoskeleton and hence it may be a way how these
rafts aggregate upon stimulation

Recruitment of cytoskeleton in response to activation of the cell to external signals

 The plasma membrane maintains
homeostasis

Plasmalemma keeps the cell contents in,

and foreign material out, and providing controlled
avenues for the transportation of fuel, fluids and
waste.

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 The membrane allows passage of beneficial
materials by the process of diffusion

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Only small, nonpolar molecules can pass through the membrane through
simple diffusion.
Plasma membranes allow water, oxygen and carbon dioxide to pass through by osmosis, or passive diffusion.
Cells must transfer essential ions and small polar molecules
across semi-permeable plasma membranes.
The cell membrane express transmembrane proteins
that provide passage to molecules that the lipid tails would
otherwise block.

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 Membrane transport

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• For polar that need to cross the plasma membrane, cells

use transport systems based on:.
• Channels open a gate for molecules to flow with their
concentration gradient.
• Transporters bond to specific types of molecules and
carry them through the membrane.
• Pumps push molecules against a concentration
gradient.
Transport Across the Membrane

Two main categories of molecule

transport exist in cells: passive transport
and active transport.

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 Passive Transport
Net movement of material from hi to low
concentration

– Diffusion
– Osmosis
– Facilitated Diffusion

concentration gradient

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 Molecule Transport
Small uncharged molecules could easily cross the
membrane via passive transport, in the form of simple
diffusion through a concentration gradient.

O2
CO2
H2O
EtOH
 Osmosis
• Refers to movement of water across a
semipermeable membrane
– Permeable to water
– Impermeable to dissolved materials
• Water always moves from
low to high solute
Hypotonic → Hypertonic

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(a) Isotonic (b) Hypertonic (c) Hypotonic
Water concentration inside the Water concentration in the Water concentration outside
cells is equal to the water surrounding medium is lower the cell is greater than it is
concentration in the fluid than it is in the cell. Water inside the cell. Water moves
surrounding the cell. Therefore, moves out of the cells and the into the cell, which swell and
there is no net movement of cells shrivel (crenate). sometimes burst – a process
water either into or out of the called lysis.
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Facilitation diffusion (catalyzed transport) – diffusion that
is assisted by proteins (channel or carrier proteins).

Sugars

These transport proteins work passively, meaning that the cell doesn't have to expend
energy sending the solute in or out.
 Transport Protein

Channel span the Carrier proteins bind to the

membrane and let solute and lead it through the
to diffuse through bilayer.
some polar and Shape change resulting from
charge molecules. solute interaction with transport
*Aquaporins let for protein.
the rapid transport
of water.
Transport Proteins are specific

glucose

Na+
Model for the facilitated diffusion of glucose

The glucose transporter alternates between two conformations,

in which a glucose-binding site is alternately exposed on the
outside and the inside of the cell.

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Affinity of glucose (or other monosaccharides) to the transporter on

both sides can be characterized by specific equilibrium constants.
Net transport is possible only in the direction of the lower
concentration of the sugar.
 Glucose transporters
14 GLUTS are encoded by human genome.
Each glucose transporter isoform plays a specific role in glucose metabolism
determined by its pattern of tissue expression, substrate specificity, transport
kinetics, and regulated expression in different physiological conditions.

GLUTs are integral membrane proteins that contain 12 membrane-spanning helices with both the
amino and carboxyl termini exposed on the cytoplasmic side of the plasma membrane.

GLUT proteins transport glucose and related hexoses according to a model of

alternate conformation.
GLUT is a type of uniporter transporter protein.
On the basis of sequence similarities, the GLUT family has been divided into a few subclasses
Name Distribution Notes

Is widely distributed in fetal tissues. Levels in cell membranes are

In the adult, it is expressed at highest levels in erythrocytes increased by reduced glucose
and also in the endothelial cells of barrier tissues such as levels and decreased by increased
GLUT1
the blood–brain barrier. glucose levels.
It is responsible for the low level of basal glucose uptake GLUT1 expression is upregulated in
required to sustain respiration in all cells. many tumors.

Is expressed by renal tubular cells, liver cells, pancreatic

beta cells and basolateral membrane of the small intestine
epithelium.
Is a bidirectional transporter, allowing glucose to flow in 2
directions.
Is a high-frequency and low-affinity
GLUT2 Bidirectionality is required in liver cells to uptake glucose
isoform.
for glycolysis, and release of glucose during
gluconeogenesis.
All three monosaccharides (glucose, galactose, and
fructose) are transported from the intestinal mucosal cell
into the portal circulation by GLUT2.
Is a high-affinity isoform, allowing it
GLUT3 Expressed mostly in neurons and in the placenta. to transport even in times of low
glucose concentrations.

Is the insulin-regulated glucose

Found in adipose tissues and striated muscle (skeletal
GLUT4 transporter; responsible for insulin-
muscle and cardiac muscle).
regulated glucose storage.

GLUT14 testicle similarity to GLUT3

 Channel proteins

Form open pores in the membrane, allowing small

molecules of the appropriate size and charge to pass
freely through the lipid bilayer

Ion channels are highly selective because narrow

pores in the channel restrict passage to ions of the
appropriate size and charge.
Thus, specific channel proteins allow the passage of Na+,
K+, Ca2+, and Cl- across the membrane.

Transport through channels is extremely rapid.

More than a million ions per second flow through open
channels—a flow rate approximately a thousand times
greater than the rate of transport by carrier proteins.

Ion channels are present in the membranes of all cells.

They have been especially well studied in nerve and muscle,
where their regulated opening and closing is responsible for the
transmission of electric signals.

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 Gated Channels
Most ion channels are not permanently open.
Ion channels are regulated by “gates” that
transiently open in response to specific
stimuli.

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 Gated Channels
Chemical
Some channels called messenger, e.g.
ligand-gated insulin, cGMP,
G proteins
channels open in
response to the
binding of
neurotransmitters or
Ca2+
other signaling
molecules.
*Ligand-gated channels are the
location where anesthetic
agents act to block the spread
of action potentials.
 Gated Channels
Voltage-gated
Voltage change channels open in
stimulus response to
changes in electric
potential across the
plasma membrane.

Na+
 Gated Channels

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• Mechanically gated channels (mechanosensitive,

MSCs) open in response to mechanical stimulus, e.g. the
tactile response of the hair cells in the inner ear.
MSCs are probably the basis of the senses of hearing and touch and sense
the stress needed for muscular coordination.
• Stretch-activated or stretch-gated ion channels are ion
channels which open their pores in response to
mechanical deformation of neuron’s plasma membrane.
The opening of these channels results in a non-specific ionic flow, which
depolarizes the afferent nerve fiber, and may produce action potentials
with sufficient depolarization.
The opening of these channels is central to a neuron’s response to
pressure, often osmotic pressure and blood pressure, to regulate ionic
flow in internal environments.
 Ungated Channel
• K+ leak channels are
the most common ion
channels.
They are ungated and
“leak” K+ the ions most
responsible for
establishing a potential
difference across the
plasmalemma.

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Facilitated diffusion can occur via ionphores
Ionophores are a lipid-soluble molecule transporting ions across
the lipid bilayer of the cell membrane.
The two broad classifications of ionophores are:
• Chemical compounds (mobile ion carriers) that bind to a particular
ion, shielding its charge from the surrounding environment, and thus
facilitating its crossing of the hydrophobic interior of the lipid
membrane;
• Channel formers that introduce a hydrophilic pore into the
membrane, allowing ions to pass through while avoiding contact
with the membrane's hydrophobic interior.
Many antibiotics, particularly the macrolide antibiotics, are
ionophores that exhibit high affinities for Na+ or K+

Gramicydn
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 Active Transport
Move materials from low to high concentration.
Requires cell to expend energy – ATP is needed to
pump the solute in or out.
Equivalent to running diffusion in reverse.

Active transport - An energy-consuming type of carrier (ATP being used) transports molecules
(usually to an area of higher concentration).
 Primary Active Transport
The membrane proteins that bind to and transport the molecules or
ions against their gradients posses the enzymatic activity to hydrolyze
or break down ATP to harness its energy.
Known as ATPase, these transporters are ATP-powered pumps that
directly hydrolyze ATP to ADP and inorganic phosphate.
 Primary Active Transport

4 classes of transport proteins function as ATP-powered

pumps to transport ions and molecules against their
concentration gradient.
All have ATP-binding sites on the cytoplasmic side of the membrane.

Class Substrate(s) transported

P - named for the phosporylation of Ions (H+, Na+, K+, Ca+2)
one of subunits transport protein
F - ATP syntase works in reverse H+ only
V - maintain the low pH of lysosomes H+ only
ABC - „ATP binding cassete” Exporting: ions, drugs, xenobiotics
superfamily
 ABC transporters
and multidrug resistance

The ABC transporters are associated with

multidrug resistance (pumping nonpolar toxic
molecules out of the cell).
The classical MDR phenotype is characterized
by a reduced ability to accumulate drugs, due to
activity of an energy-dependent uni-directional,
membrane-bound, drug-efflux pump with broad
substrate specificity.
MDR is major limitation in cancer therapy.

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 ABC transporters
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These pumps can move substrates in (influx) or out (efflux) of cells.

The human genome contains 49 ABC genes, arranged in 8 subfamilies.
ABC transporters are expressed predominantly in the liver, intestine,
blood-brain barrier, blood-testis barrier, placenta and kidney.

ABC transporters participate in the movement of most drugs and their

metabolites across cell surface and cellular organelle membranes.
Defects in these genes can be important in terms of cancer
therapy, pharmacokinetics and innumerable pharmacogenetic
disorders.
Mutations in at least 11 of ABC genes are already known to cause severe
inherited diseases (eg cystic fibrosis and X-linked adrenoleukodystrophy [X-
ALD]).
 Active Transport
The sodium-potassium pump in conjunction with the potassium leak
channel, allows the cell to control it's membrane potential.
The sodium-potassium-ATPase pumps sodium out and potassium in,
which creates a high concentration of potassium inside the cell, and a low
concentration outside.
The potassium leak channel allows the potassium to leak out, which gives
the cell a negative charge on the inside.

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Ion gradients across the plasma membrane of
a typical mammalian cell

The low concentrations of Na+ and Cl- balance the high intracellular
concentration of organic compounds, equalizing the osmotic
pressure and preventing the net influx of water.

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 Membrane potential
Differences in ions’ concentration on opposite sides of a cellular membrane lead
to a voltage called the membrane potential (or transmembrane potential or
membrane voltage).
It is the separation of these charges across the membrane that is the basis
of the membrane voltage.
Changes in membrane potential elicit action potentials and give cells the
ability to send messages around the body.

www.cetbiology.com
 Secondary Active Transport

The process by which ion gradients generated by ATP-powered

pumps are used to power transport of other molecules and ions
against their own concentration gradients.

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Active secondary transport driven by the Na+ gradient
is responsible for the uptake of glucose from the intestinal lumen.
The transporter coordinately binds and transports 1 glucose and 2 Na+
into the cell.
The transport of Na+ in the energetically favorable direction drives the
uptake of glucose against its concentration gradient.

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 Transport of molecules
Uniports simply move solutes from one side to another.
Cotransport systems work by simultaneously sending two solutes
across the lipid bilayer.
There are two types of cotransport systems:
• symport, in which the solutes are sent in the same direction,
• antiport, in which they are sent in opposite directions.

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 Drug Transport
Most drugs must be transferred from the side of administration to the
blood stream in order to reach their target tissues within the body.

www.publications.nigms.nih.gov

www.pharmlabs.unc.edu www.physics.cancer.gov

Intravenously administered drugs are Orally administrated drugs must be dissolve in

completely absorbed and the total dose gastrointestinal fluid and then penetrate the
of drugs reaches the circulation. epithelial cells of the intestinal mucosa in order
to enter the circulation.
 www.pharm.sdu.edu.cn

Drug Transport
Many drug transporters function as primary and secondary active
transporters.
The structures, function and tissue distribution vary widely.
Drug transporters Transport

Solute carriers (SLC’s): Secondary active transort

•PEPT Proton (H+) and di- and tri-peptide cotransport

Peptide transporters B-lactam antibiotics (penicilin), ACE (angitenisn converting enzyme) inhibitors

•OATP Amphipathic organic coumpounds such as:

Organic anion transporting bile salts, steroids, thyroid hormones
polypeptides

•Organic ions transporters Organic ions

Metformin – drug to treat type 2 diabetes

•H+/organic cation antiporters Organic cations are excreted and H+ are taken up

ABC transporters Primary active transport

Export of ions, drugs and xenobiotics
anticancer-, antiviral-, immunosuppressive agents, calcium channels blockers
 Drug Transport
The blood–brain barrier (BBB) is a highly selective
permeability barrier that separates the circulating blood
from the brain extracellular fluid in the central nervous
system.
The blood–brain barrier is formed by capillary
endothelial cells, which are connected by tight
junctions.
The functional consequence of these features is that
brain capillaries act in a passive manner to restrict
penetration of hydrophilic or large substances.

The blood–brain barrier allows the passage of water,

some gases, and lipid soluble molecules by passive
diffusion, as well as the selective transport of
molecules such as glucose and amino acids that are
crucial to neural function.
Highly lipophilic drugs (like most antiepileptic drugs)
www.jpet.aspetjournals.org
penetrate easily through the BBB by simple diffusion.
 Macromolecules Transport

To transport the macromolecules (proteins,

polynucleotides, and polysaccharides), cells rely on active
transport.
The active transport of macromolecules involved:
• exocytosis and
• endocytosis.

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 Membrane transport
of macromolecules
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In exocytosis the contents of special vesicles are

released when the vesicle fuses with the cell
membrane.

In endocytosis the membrane depresses and pinches

off, enclosing the molecule.
Two different sizes are formed – pinocytotic (small)
and phagocytic (large).
 Endocytosis
Endocytosis is a kind of bulk transport where large amounts of
material are brought into a cell.

The receptors play a role in identifying what is "good to eat" and that
membrane is removed from the plasma membrane.
 Receptor-mediated endocytosis
The macromolecules to be internalized first bind to specific cell surface
receptors, allowing the cell to select what molecules to take and what to reject.
These receptors are concentrated in specialized regions of the plasma
membrane, called clathrin-coated pits.
The pits bud from the membrane to form small clathrin-coated vesicles
containing the receptors and their bound macromolecules (ligands).

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 Endocytosis
The molecules that have been ingested are then deposited at the cytosol,
where the cell uses them and most of the vesicle is reintegrated into the cellular
membrane.
The clathrin-coated vesicles fuse with early endosomes, in which their
contents are sorted for transport to lysosomes or recycling to the plasma
membrane.
In some cases the vesicle bypasses the secondary lysosome and goes directly to
its target.
Recycling to the plasma membrane

Many receptors (like the LDL receptor)

being returned to the plasma
membrane following dissociation of
their bound ligands in early
endosomes.
The recycling of the receptors
results in the continuous
internalization of their ligands.
Each LDL receptor, for example, makes a round-
trip from the plasma membrane to endosomes
and back approximately every 10 minutes.
About 50% of the plasma membrane
is internalized by receptor-mediated
endocytosis every hour and must
therefore be replaced at an equivalent
rate.
Most of this replacement is the result
of receptor recycling; only about 5% of
the cell surface is newly synthesized
per hour.
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 Exocytosis

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Exocytosis is a kind of bulk transport, in which large amounts of

material are moved out of a cell.
Macromolecules are packaged in a vesicle that separates them out
from the rest of the cell.
The vesicle fuses with its specific membrane structure and its
contents is released.
A new membrane is added to the plasma membrane (from the
secretion vesicle).
 Exocytosis
Proteins, that are to be secreted from the cell are made on ER.
They are then transported to the Golgi complex by ER induced
vesicles.
The Golgi complex sorts and packages the proteins into vesicles
that separate themselves off the Golgi complex and eventually
fuse with the cellular membrane.

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 Exocytosis
Some molecules are secreted continually from the cell,
but others are selectively secreted.
To control secretion, specific substances are stored in secretory vesicles,
which are released when triggered by an extracellular signal (hormone).
The signal triggers exocytosis, causing the secretory vesicles to fuse with
the cellular membrane, releasing the substances outside the cell.

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