Membrane Proteins

Membrane Proteins
• Proteins that interact with, or are part of, biological
membranes.

• Membrane proteins may be permanently anchored

in the membrane or are part of the membrane

• Membrane proteins may only be temporarily

attached to the lipid bilayer or to other proteins.
 Membrane Proteins
• Membrane proteins may be classified as:

1. Integral (intrinsic)

2. Peripheral (extrinsic)

3. Having a lipid anchor

peripheral
lipid
anchor

lipid bilayer

Membrane
integral Proteins
 Membrane Proteins
• Membrane proteins are a common type of proteins
along with:
– Soluble globular proteins
– Fibrous proteins
– Disordered proteins

• They are targets of over 50% of all modern medicinal

drugs

• It is estimated that 20–30% of all genes in

most genomes encode membrane proteins.
 Functions of Membrane Proteins
• Membrane proteins perform a variety of functions vital
to the survival of organisms

• Membrane receptor proteins relay signals between the

cell's internal and external environments.

• Transport proteins move molecules and ions across the

membrane.
– They can be categorized according to the Transporter
Classification database.
 Functions of Membrane Proteins

• Membrane enzymes may have many activities,

such as oxidoreductase, transferase or
hydrolase.

• Cell adhesion molecules allow cells to identify

each other and interact.
– For example, proteins involved in immune
response.
 Functions of Membrane Proteins
• Cell surface identity markers as proteins expressed
on the surface of cells that often conveniently serve
as markers of specific cell types.

• Attachments to the cytoskeleton to provide the cell

with structure and support
Plasma Membrane Proteins

PROTEINS CAN
MOVE IN THE
MEMBRANE,
TOO!
 Membrane Proteins
• Membrane proteins can be made of alpha helices or
beta strands, or the combination of both alpha helices
or beta strands.

• Membrane-spanning alpha helices are the most

common structural motif in membrane proteins.

• Eg. The enzyme protein called prostaglandin is made

entirely of the alpha helices.

• Eg. The channel protein called porin is made up of

entirely beta strands
Prostaglandin
 Membrane Proteins
• Examination of the primary structure reveals that most
amino acids in the membrane proteins are nonpolar and
very few are charged.

• One of the first alpha - proteins found was the

bacteriorhodopsin.

• It uses light energy to transport protons from inside the cell

to outside generating a proton gradient used to form ATP.

• Voltage-gated ion channels, such as potassium and chloride

channels, are examples of alpha-helical transmembrane
proteins.
Bacteriorhodopsin
Nonpolar regions of the protein are embedded in
the interior of the bilayer

Polar regions of the protein protrude from both

sides of the bilayer
 Membrane Proteins
• Beta strands form channel proteins.

• They are less common than alpha helices.

• Each strand is hydrogen bonded to its neighbour in an

anti-parallel arrangement, forming a single beta sheet.

• The beta sheet then curls up to form a hollow cylinder

that forms a channel in the membrane.
 Membrane Proteins
• The outside surface of the β-sheet is non-polar and
interacts with the hydrocarbon core of the membrane

• The inside channel is hydrophilic and filled with water.

• The arrangement of polar and non-polar is

accomplished by the alternation of hydrophobic and
hydrophilic amino acids along each β strand.
Porins
 Membrane Proteins
• Many membrane proteins have quaternary structures
consisting of multiple subunits.

• This oligomerization in membrane proteins is beneficial

to their functions, stability, genetic efficiency

• May optimize productive output per unit area of the

membrane.
 Membrane Proteins
• Usually consist of two subunits which are connected by
a bridge so that electrons can be transferred between
them.

• Van der Waals interactions hold the tertiary and

quaternary structures together in the transmembrane
region.

• These interactions allow for flexibility in the structure

to accommodate for necessary functions.
1. Integral Membrane Proteins
 Integral Membrane Proteins
• Characterized by strong interaction with the membrane

• Permanently attached to the membrane.

• Can be separated from the membranes only using

detergents, nonpolar solvents, or sometimes
denaturing agents.

• Most integral proteins contain residues with hydrophobic

side chains
– Interact with fatty acyl groups of the membrane phospholipids,
thus anchoring the protein to the membrane
 Integral Membrane Proteins
• Can be classified according to their relationship
with the bilayer

• The integral membrane proteins are classified as:

a) Integral polytopic proteins (Transmembrane

proteins) that span across the membrane

b) Integral monotopic proteins that are attached to only

one side of the membrane.
 Integral Polytopic Proteins
• Also known as transmembrane proteins, are integral
membrane proteins that span across the membrane.

• They are the most common among integral proteins.

• They may cross the membrane only once or several times,

weaving in and out.

• These proteins may have different transmembrane topology

 Integral Polytopic Proteins

• Transmembrane proteins have one of two

structural architectures:

– α helix bundle proteins: present in all types

of biological membranes

– β barrel proteins: found only in outer membranes of G-

bacteria, lipid-rich cell walls of
a few G+, and outer membranes of mitochondria
and chloroplasts.
Integral Polytopic Proteins
 Integral Monotopic Proteins
• Integral membrane proteins that are attached to only
one side of the membrane

• Do not span the whole way across the bilayer

• These proteins are permanently bounded to the

membrane but only from one side.

• Many of these proteins are enzymes.

– E.g cyclooxygenase and carnitine O-palmitoyltransferase.
 Integral Monotopic Proteins
• Cyclooxygenase is involved in the formation of
prostanoids.

• Anti-inflammatory drugs, such as aspirin and ibuprofen,

work to relieve symptoms of inflammation and pain by
inhibiting this enzyme.

• Carnitine O-palmitoyltransferase is a mitochondria

transferase enzyme: participates in the metabolism of
palmitoylcarnitine into palmitoyl-CoA.
2. Peripheral Membrane Proteins
 Peripheral Membrane Proteins
• Have a much weaker interaction with the membrane than
integral proteins

• Temporarily attached either to the lipid bilayer or to

integral proteins

• By a combination of hydrophobic, electrostatic, and other

non-covalent interactions.

• Do not interact with the hydrophobic core of the

phospholipid bilayer.
 Peripheral Membrane Proteins
• Peripheral proteins dissociate following
treatment with a polar reagent
– E.g a solution with an elevated pH or high salt
concentrations.
 Membrane Proteins
• Integral and peripheral proteins may be post-
translationally modified with added:
– Fatty acid,
– Prenyl chains,
– GPI (glycosylphosphatidylinositol)

• Which may be anchored in the lipid bilayer.

 Polypeptide Toxins
• Polypeptide toxins and many antibacterial peptides
such as: colicins, hemolysins and certain apoptosis
proteins

• Sometimes considered a separate category.

• These proteins are water-soluble but can aggregate

and associate irreversibly with the lipid bilayer and
become reversibly or irreversibly membrane-
associated.
Hemolysin
 Transport Proteins
• Due to the nature of the lipid bilayer, many molecules
cannot enter or exit the cell because of size or charge.

• Membrane proteins function to assist in the

transportation of such molecules across the lipid bilayer.

• Trans-membrane proteins participate in either passive or

active transport.
(1) A single a helix
(2) As multiple a helices
(3) As a rolled-up b sheet (a barrel). Some of these "single-pass" and "multipass" proteins have a
covalently attached fatty acid chain inserted in the cytosolic lipid monolayer. Other membrane proteins
are exposed at only one side of the membrane.
(4) Some of these are anchored to the cytosolic surface by an amphipathic a helix that partitions into the
cytosolic monolayer of the lipid bilayer through the hydrophobic face of the helix.
(5) Others are attached to the bilayer solely by a covalently attached lipid chain either a fatty acid chain or
a prenyl group in the cytosolic monolayer or,
(6) Via an oligosaccharide linker, to phosphatidylinositol in the non cytosolic monolayer.
(7, 8) Finally, many proteins are attached to the membrane only by non-covalent interactions with other
membrane proteins.
3. Lipid Anchor
 Lipid Anchor
• Some proteins bind to membranes via a covalently attached
lipid anchor, that inserts into the bilayer.

• A protein may link to the cytosolic surface of the plasma

membrane via a covalently attached fatty acid (e.g., palmitate
or myristate) or an isoprenoid group.

• Palmitate is usually attached via an ester linkage to the thiol

of a cysteine residue.

• A protein may be released from plasma membrane to cytosol

via depalmitoylation, hydrolysis of the ester link.
 Lipid Anchor

lipid
anchor O O C

H3C (CH2)14 C S CH2 CH cysteine

residue
NH
membrane palmitate
 Glycoproteins
• Proteins that contain oligosaccharide chains
(glycans) covalently attached to polypeptide side-chains.

• The carbohydrate is attached to the protein in

a cotranslational or posttranslational modification.

• This process is known as glycosylation.

• Secreted extracellular proteins are often glycosylated.

 Cells are sugar coated
 Sugar moieties protruding outward in plasma
membrane.
 No sugar on inside
 Types of glycosylation
1. N-glycosylation: sugars are attached to nitrogen, typically
on the amide side-chain of asparagine.

2. O-glycosylation: sugars are attached to oxygen, typically

on serine or threonine but also on non-canonical amino
acids such as hydroxylysine & hydroxyproline.

3. P-glycosylation: sugars are attached to phosphorus on

a phosphoserine.
 Types of glycosylation
4. C-glycosylation: sugars are attached directly to
carbon, such as in the addition of
mannose to tryptophan.

5. Glypiation: a GPI glycolipid is attached to the C-

terminus of a polypeptide, serving as a membrane
anchor.
Biological Membranes Summary