Chapter 8 S1 1819 Cellular Membranes Part 1 PDF

Cellular Membranes
© 2013 John Wiley & Sons, Inc. All rights reserved.

1
Keys
• Describe the functions of cellular membranes.

• Elucidate the chemical components and properties of cell membranes.
• Describe the development of the models to the Fluid-Mosaic Model.
• Explain the role of carbohydrates in membrane structure.
• Describe the types of proteins found in membranes and their roles.
• Stress the importance and detection of membrane fluidity in living cells.
• Describe biological membrane asymmetry.
• Describe the mechanisms to transport materials across membranes:
simple and facilitated diffusion, channel proteins, active transport.
• Explain the process involved in generating an action potential and
propagating the signal across the synapse to the postsynaptic cell.

2
Introduction
The trilaminar appearance of membranes as

revealed by electron micrograph of the plasma
membrane and sarcoplasmic reticulum.
• Plasma membrane: The outer boundary of the cell that separates it

from the world is a thin, fragile structure about 5 – 10 nm thick.
• Not detectable with light microscope need electron microscope.
• The 2 dark-staining layers in the electron micrographs correspond
primarily to the inner & outer polar surfaces of the bilayer
• All membranes examined closely (plasma, nuclear or cytoplasmic) from
plants, animals or microorganisms have the same ultrastructure

3
An Overview of Membrane Functions
• Compartmentalization (1)
Membranes form continuous sheets
that enclose intracellular
compartments.
• Scaffold for biochemical activities (2)
Membranes provide a framework
that organizes enzymes for effective
interaction.
• Selectively permeable barrier (3)
Membranes allow regulated
exchange of substances between
compartments.
A summary of membrane
functions in a plant cell.

4
An Overview of Membrane Functions
• Transporting solutes (4)
Membrane proteins facilitate the
movement of substances between
compartments.
• Responding to external signals (5)
Membrane receptors transduce
signals from outside the cell in
response to specific ligands.
• Intracellular interaction (6)
Membranes mediate recognition
and interaction between adjacent
cells.
• Energy transduction (7)
Membranes transduce
photosynthetic energy, convert
chemical energy to ATP, and store A summary of membrane
energy. functions in a plant cell.

5
A Brief History of Studies on Plasma
Membrane Structure
Calculating the
surface area of a
lipid preparation
• Membranes were found to be mostly composed of lipids because their

dissolving power matched that of oil.
• Gorter and Grendel (1925): first proposed lipid bilayer
– extracted lipids from red blood cell (RBC) membrane, measured their surface area on
H2O & compared it to estimated RBC surface area
– The lipid bilayer accounted for the 2:1 ratio of lipid to cell surface area (used RBC)
– Plasma membrane is the only lipid-containing structure in cell.
– Propose lipid bilayer (bimolecular layer of lipids) with hydrophilic (Polar) heads
pointed out on both sides & hydrophobic fatty acid (acyl) tails protected from
aqueous environment.
6
Membrane Structure
Early models
representing
the lipid
bilayer
• 1920s & 1930s - evidence accrued that there must be more to

membranes than lipid bilayer.
• Hugh Davson & James Danielli (1935) - proposed that membrane was
composed of lipid bilayer lined on inner & outer surfaces by layer of
globular proteins.

7
Membrane Structure
Early models
representing
the lipid
bilayer
• In mid to late 1950s, with techniques for preparing & staining tissue, J.
D. Robertson & others were able to resolve cell membranes in the
electron microscope.
• D. Robertson proposed the Unit Membrane Model - He suggested that
all membranes were the same.
• This hypothesis was rejected because a number of variations between
different membranes was catalogued.

8
Membrane Structure
S. Jonathan Singer & Garth Nicolson (1972) - proposed the
Fluid-mosaic model
9
Membrane Structure
• The fluid-mosaic model

• Core lipid bilayer exists in a fluid state, capable of movement.
• Membrane proteins form a mosaic of particles penetrating the lipids.

10
The fluid-mosaic model
Molecular model of
the membrane of a
synaptic vesicle
constructed with
various proteins
embedded into the
lipid bilayer
A. Lipid bilayer remains core of membrane, but it is not frozen &

immobile, but fluid; individual lipids move laterally within plane of
membrane
B. Proteins distributed differently - mosaic of discontinuous particles that
penetrate into or through membrane or contact its polar heads
without penetrating the membrane

11
The Chemical Composition of Membranes
• Membrane composition
– The lipid and protein components are
bound together by non-covalent bonds.
– Membranes also contain
carbohydrates.
• Membrane lipids are amphipathic with three main

types:
• Phosphoglycerides are diacylglycerides with small functional head
groups linked to the glycerol backbone by phosphate ester bonds.
• Sphingolipids are ceramides formed by the attachment of
sphingosine to fatty acids.
• Cholesterol is a smaller and less amphipathic lipid that is only found
in animals. 12
Chemical structure of membrane lipids
Polar P
Glycerol
Fatty acid
• Phosphoglycerides are chain
diacylglycerides with
small functional head
groups linked to the
glycerol backbone by The chemical structure
phosphate ester bonds. of membrane lipids
• Sphingolipids are
ceramides formed by the
attachment of
sphingosine to fatty acids.
• Cholesterol is a smaller
and less amphipathic lipid
that is only found in
animals.

13
Sphingolipids
• A non-glycerol based polar lipids
• The overall shape is similar to phospholipids
but have different structure called
sphingosine.
• Sphingosine is an alcohol amine lipid with a
long hydrocarbon chain.
• They have a single unsaturated near the polar
end.
Prepared by: Dr. A. Ameer Allaith

Sphingosine
• The basic unit for sphingolipids. It is C18
amino alcohol. It has two functional groups:
amino group and hydroxyl groups
Modified from Dr. A. Ameer Allaith

Sphingolipids
http://www.wikiwand.com/en/Sphingolipid
16
Chemical structure of membrane lipids
• Cholesterol is a smaller and
less amphipathic lipid that is
only found in animals.
• It is missing from most plant
& all bacteria cell membranes
• Small hydrophilic hydroxyl
group is oriented toward
membrane surface; the rest is
embedded in the lipid bilayer
• Carbon rings are flat and rigid;
interfere with movement of
phospholipid fatty acid tails
Cholesterol molecules (green) oriented with
their small hydrophilic end facing the external
surface of the bilayer

17
18
19
Liposomes: synthetic vesicles
• The Nature and Importance of the Lipid Bilayer

– Membrane lipid composition is characteristic of specific membranes.
– Lipids give membranes the ability to fuse, form networks, and separate
charge.
– Lipid bilayers assemble spontaneously in aqueous solutions as in liposomes.

20
The dynamic properties of plasma membranes
Division: invagination of
Movement: ruffling of Fusion: plasma
the plasma membrane
the plasma membrane membranes of sperm and
towards the cell center
of a migrating cell egg unite
during cell division

21
• The Asymmetry of
Membrane Lipids
– Inner and outer membrane
leaflets have different lipid
compositions.
– Provides different physico-
chemical properties
appropriate for different
interactions
– Membrane lipids move
easily within a leaflet but
only rarely “flip-flop”
SM: sphingomyelin PE: phosphatidylethanolamine
PC: phosphatidylcholine PI: phosphatidylinositol
PS: phosphatidylserine Cl: cholesterol

22
Membrane Carbohydrates
– Eukaryotic cell plasma membranes also contain
carbohydrate.
– All membrane carbohydrates face toward outside of
cells into extracellular space or toward organelle
interior (carbohydrates of internal cellular
membranes); in both cases, they face away from
cytosol
– Membranes contain carbohydrates covalently linked
to lipids and proteins on the extracellular surface of
the bilayer.

23
Two types of
linkages that join
sugars to a
polypeptide chain
Blood-group
antigens
– Glycoproteins have short (oligosaccharides with < ~15 sugars per
chain), branched carbohydrates for interactions with other cells and
structures outside the cell.
– Glycolipids have short, branched oligosaccharides chains (Larger) that
may be cell-to-cell recognition sites.

24
Blood-group
antigens
On RBCs, glycolipids determine ABO blood type
1. Person with blood type A has enzyme that adds N-acetylgalactosamine to end
of chain
2. Person with blood type B has enzyme that adds galactose to chain terminus
3. AB people possess both enzymes;
4. people with type O blood lack enzymes capable of attaching either terminal
sugar
25
The Structure and Functions of Membrane
Proteins
• Membrane proteins attach to the bilayer
asymmetrically, giving the membrane a
distinct “sidedness”
• Membrane proteins can be grouped into
three distinct classes:
1. Integral proteins
2. Peripheral proteins
3. Lipid-anchored membrane
proteins
Integral proteins

26
The Structure and Functions of
Membrane Proteins
Solubilization of membrane An integral protein as it resides

proteins with detergents within the plasma membrane
Membrane Protein Sidedness

• Refer to the non-uniform distribution of protein between
the outer and inner surface of the bilayer.
Text: Dr. AbdulAmeer Al-Laith

27
The Structure and Functions of Membrane
Proteins
• Penetrate and pass through lipid bilayer.
they pass entirely through bilayer
(transmembrane)
• Make up 20 -30% of all encoded proteins
• Are amphipathic, with hydrophobic
domains anchoring them in the bilayer
(hydrophobic parts contact fatty acids in
bilayer) and hydrophilic regions forming
functional domains outside of the bilayer. Integral proteins
• Hydrophilic domains protrude from both

sides of membrane (extracellular &
cytoplasmic)
• Some have only one membrane-spanning
segment; others are multispanning. 28
1.Integral proteins
Most integral membrane proteins function in the following

capacities:
A. As receptors that bind specific substances at the
membrane surface
B. As channels or transporters involved in the
movement of ions & solutes across the membrane or
C. As agents that transfer electrons during the
processes of photosynthesis & respiration

29
Membrane Proteins
Driven by van der Waals forces between amino

acids and lipids, proteins can be surrounded by
a closely applied shell of lipid molecules.
• Channel proteins have hydrophilic cores that form
aqueous channels in the membrane-spanning region.

30
Membrane Proteins
– Are attached to the membrane by
weak bonds and are easily
solubilized.
– Located entirely outside of bilayer
on either the extracellular or
cytoplasmic side; associated with
membrane surface by
Peripheral proteins
noncovalent bonds

31
Membrane Proteins
• Best-studied peripheral proteins are
located on cytosolic membrane
surface
• They form fibrillar network that acts
as membrane skeleton
A. These proteins give mechanical support to
membrane & function as an anchor for integral
proteins
B. Other peripheral proteins on internal Peripheral proteins
membrane surface function as enzymes,
specialized coats or factors that transmit
transmembrane signals
32
Membrane Proteins
proteins
– located outside bilayer on either
extracellular or cytoplasmic side
– They are covalently linked to
membrane lipid situated within
bilayer
Lipid-anchored proteins

33
Membrane Proteins
This is an
oligosaccharide part
linking a protein to lipid
An extracellular
protein
A cytoplsmic proten Lipid-anchored proteins

directly linked to
membrane lipid 34
Modified From: Dr. A. Ameer Allaith © 2013 John Wiley & Sons, Inc. All rights reserved.
Membrane Proteins
proteins
Kinds of lipid anchor and their orientation:
– Linked via a short oligosaccharide to a
phospholipid, external face.
Glycophosphatidylinositol (GPI)-linked
proteins found on the outer leaflet
can be released by inositol-specific
phospholipases.
Lipid-anchored proteins
– Some inner-leaflet proteins are
anchored to membrane lipids by long
hydrocarbon chains.

35
Membrane Proteins
Ectoplasmic
vs
protoplasmic
• Distribution of Integral Proteins: Freeze-Fracture Analysis

– Freeze-fracture technique divides the phospholipid
leaflets of the membrane.
– Integral membrane proteins appear as bumps and pits
using the electron microscope.
– The heterogeneity of protein distribution is shown.

36
Identification of Transmembrane
Domains
• Transmembrane domain is
predicated from amino acid
sequence (deduced from
the gene)
• These segments typically

consists of 20-30 amino
Glycophorin A, an integral protein
with a single transmembrane domain acids mostly non-polar, and
with a Gly-X-X-X-Gly sequence have -helical secondary
structure.

37
Membrane Lipids and Membrane
Fluidity
Physical state of membrane lipids described by
fluidity (or viscosity)
– Fluidity: measure of ease of flow

– Viscosity: measure of resistance to flow
Lipids exist in 2 states

– Solid
– Liquid phase of varying viscosity depending on
temperature.

38
Membrane Lipids and Membrane
Fluidity
• Membrane lipids exist in gel or liquid-crystal phases
depending on:
– temperature,
– lipid composition
– saturation
– the presence of cholesterol.
• Liquid-crystal membranes predominate

• Unsaturated fatty acids lower the temperature at which
the liquid-crystal/gel phase transition occurs (transition
temperature).
• Transition temperature - temperature at which
membrane goes from fluid state to crystalline gel
39
Factors affecting membrane fluidity:1
1. Temperature
2. Degree of saturation
3. Length of Fatty acyl chains.
4. Amount of cholesterol.
Effect of Temperature
• At warm temperature the bilayer is a liquid-
crystal state (2D). Molecules have specified
orientations and can rotate.
• When temperature is lower, it will change to a
solid-like (gel) state at transition temperature.
Movement of molecules is restricted.
Membrane Lipids and Membrane Fluidity
Structure depends on the temperature
Structure of the lipid bilayer depends on the temperature:

above and below the transition temperature.
• The Importance of Membrane Fluidity

– The fluidity of membranes is a compromise between structural rigidity and
complete fluidity.
– Membrane fluidity makes it possible for proteins to move in the membrane
and for membranes to assemble and grow.
42
Membrane Lipids and Membrane Fluidity
Structure depends on the temperature
Structure of the
lipid bilayer
depends on the
temperature:
above and below
the transition
temperature.
• Maintaining Membrane Fluidity

– Organisms (other than birds and mammals) maintain membrane
fluidity as temperature changes by altering the composition of
membrane lipids.
– Remodeling lipid bilayers involves saturation or desaturation of acyl
chains and replacement of acyl chains by phospholipases or
acyltransferases.
43
Degree of unsaturation
• The greater the degree of unsaturation of the
fatty acids, the lower the temperature of the
melting point.
• This means that more unsaturation results in
increasing the fluidity, and vise versa.
• Unsaturation produces bent and interferes
with the packing of the chains.
45
Length of Fatty acid chain
• The shorter the fatty acyl chain of

phospholipid, the lower its melting point.
• This means that longer FA chains decrease
fluidity, while shorter FA chains increase it.
Prepared By: Dr. AbdulAmeer Al-Laith

Effects of cholesterol on Fluidity
Cholesterol – affects membrane physical state; interacts with

membrane phospholipid fatty acid chains & alters the way the
fatty acids pack together
1. Because of their orientation within the bilayer, cholesterol
disrupts the close packing of fatty acyl chains & interferes
with their mobility.
2. It tends to abolish sharp transition temperatures & creates
a condition of intermediate fluidity
3. In physiological terms, it tends to raise membrane
durability & lower membrane permeability
Importance of membrane fluidity
A. Membrane fluidity provides a perfect compromise

between rigid, ordered structure & a totally fluid,
nonviscous liquid:
– In the rigid structure - mobility would be absent
– In the totally fluid structure - components could not be
oriented; structural organization & mechanical support
would be lacking

48
B. Moderate fluidity also allows interactions to take
place within membrane; clusters of membrane
proteins can assemble at particular sites within
membrane & form specialized structures
– Among the specialized structures: intercellular junctions,
light-capturing photosynthetic complexes
– Molecules that interact can come together, carry out
necessary reaction & move apart

49
C. Membranes arise only from preexisting
membranes – their growth is accomplished by
the insertion of lipid & protein components into
the fluid matrix of the membranous sheet.
D. Many of the most basic cell processes (cell

movement, growth & division; intercellular
junction formation, secretion,
endocytosis/exocytosis) depend on membrane
component movement
– These processes would probably not be possible if
membranes were rigid, nonfluid structures
50
(8.6) The Dynamic Nature of the
Plasma Membrane
• Lipid bilayer can exist in a relatively

fluid state.
• A phospholipid can move laterally
within the same leaflet with
considerable ease.
• In contrast, it takes a phospholipid
molecule a matter of hours to days to
move across to the other leaflet (flip-
flop).
• The hydrophilic head group of the
lipid must pass through the internal
hydrophobic sheet of the membrane, The possible movements of
which is thermodynamically phospholipids in a membrane
unfavorable.
51
The Dynamic Nature of the Plasma Membrane
Cell fusion to reveal mobility of membrane proteins:

fusion of human and mouse cells
• The Diffusion of Membrane Proteins after Cell Fusion

– Cell fusion is a technique whereby two different types of cells, or cells
from two different species, can be fused to produce one cell with a
common cytoplasm and a single, continuous plasma membrane.
– Cell fusion be induced by certain viruses, or with polyethylene glycol.
– Labeled proteins have shown that membrane proteins can move
between fused cell.
52
Protein Mobility – Cell Fusion 2
Findings:
• Distribution of each type of protein into the other half
after fusion increases with time. After 40 min full
random distribution is completed.
• At temperature below 15C, fusion is retarded, hence
distribution did not occur. Sharp fusion (mosaic) occurs
above 15C.
Conclusions:
• Integral proteins can move within the PM bilayer.
• This is an indication that the lipid bilayer is not rigid,
but in a dynamic state.
• Movement of bilayer is affected by temperature.

53
Measuring the diffusion
rates of membrane
• Restrictions on Protein proteins by FRAP:
and Lipid Mobility variable nature of
– Proteins can be labeled fluorescence recovery is
and tracked by dependent upon the
fluorescence recovery protein examined
after photobleaching
(FRAP) and single
particle tracking (SPT).
– Proteins can be
immobile, mobile in a
directed manner, or
exhibit random
movement.

54
Measuring the diffusion
rates of membrane
Findings: proteins by FRAP:
• Random movement of proteins variable nature of
will be seen, and the spot will fluorescence recovery is
gradually be filled with the dye. dependent upon the
The rate of recovery (the
diffusion rate) can be monitored. protein examined
This rate depends on the
nature/type of proteins.
Different proteins have different
diffusion rate.
Conclusions:
• PM Proteins can move, their rate
of diffusion vary.
• FRAP can measure the
movement of large labeled
population of proteins.

55
Control of Membrane Protein
Mobility
– Protein movements are slower than
predicted by protein size and
membrane viscosity.
– Protein movements are limited by
interactions with the cytoskeleton,
other proteins, and extracellular
materials.
– Techniques that can drag tagged
proteins within the membrane,
indicate that some proteins have
barriers to lateral diffusion.
– Genetically modified proteins Patterns of movement of
missing either intracellular or integral membrane proteins
extracellular domains are less
restricted.

56
Patterns of Movement of Integral
membrane Proteins
Findings and Conclusions:
Six types of proteins movements
were revealed by this technique.
These include:
1. Random movement (diffusion)
in PM; protein A. Rate varies
with type of proteins and the
properties of PM.
2. Immobilized (non moving)
proteins due to interaction with
underlying membrane skeleton.
Protein B
3. Movement with highly directed
manner, in a specific direction,
due to interaction with motor
protein (in the cytoplasmic side).
Protein C. Patterns of movement of
integral membrane proteins

57
Patterns of Movement of Integral
membrane Proteins
Findings and Conclusions:
Six types of proteins movements
were revealed by this technique.
These include:
4. Restricted movement/diffusion;
due to interaction with other
integral proteins. Protein D.
5. Restricted movement/diffusion
due to the fence, barriers of
proteins of PM skeleton. Protein
E
6. Restrained movement/diffusion
due to the presence of extra-
cellular materials. Protein F.
Patterns of movement of
integral membrane proteins

58
Membrane Domains and

Cell Polarity
– Differences in protein
distribution are
evident in cells of
organized tissues.
– In epithelia, the
proteins of the apical
membrane are distinct
from those of the
lateral and basal
membranes
Differentiated functions of the plasma
membrane of an epithelial cell.

59
Membrane Domains and

Cell Polarity
– Highly differentiated
sperm have a head,
midpiece, and tail that
is covered by a
continuous
membrane. Can
distinguish these
regions with antibody
staining.
Differentiation of the mammalian sperm plasma

membrane as revealed by fluorescent antibodies.

60
Copyright 2013 John Wiley & Sons, Inc.
All rights reserved. Reproduction or translation of this work
beyond that permitted in section 117 of the 1976 United
States Copyright Act without express permission of the
copyright owner is unlawful. Request for further information
should be addressed to the Permissions Department, John
Wiley & Sons, Inc. The purchaser may make back-up copies
for his/her own use only and not for distribution or resale.
The Publisher assumes no responsibility for errors,
omissions, or damages caused by the use of these programs
or from the use of the information herein.

61

Chapter 8 S1 1819 Cellular Membranes Part 1 PDF

Caricato da

Informazioni sul documento

Titolo originale

Copyright

Formati disponibili

Condividi questo documento

Condividi o incorpora il documento

Opzioni di condivisione

Hai trovato utile questo documento?

Questo contenuto è inappropriato?

Copyright:

Formati disponibili

Chapter 8 S1 1819 Cellular Membranes Part 1 PDF

Caricato da

Copyright:

Formati disponibili

Cellular Membranes

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Describe the functions of cellular membranes.

© 2013 John Wiley & Sons, Inc. All rights reserved.

The trilaminar appearance of membranes as

• Plasma membrane: The outer boundary of the cell that separates it

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Membranes were found to be mostly composed of lipids because their

• 1920s & 1930s - evidence accrued that there must be more to

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

S. Jonathan Singer & Garth Nicolson (1972) - proposed the

• The fluid-mosaic model

© 2013 John Wiley & Sons, Inc. All rights reserved.

A. Lipid bilayer remains core of membrane, but it is not frozen &

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Membrane lipids are amphipathic with three main

© 2013 John Wiley & Sons, Inc. All rights reserved.

Prepared by: Dr. A. Ameer Allaith

Modified from Dr. A. Ameer Allaith

© 2013 John Wiley & Sons, Inc. All rights reserved.

Liposomes: synthetic vesicles

• The Nature and Importance of the Lipid Bilayer

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

Solubilization of membrane An integral protein as it resides

Membrane Protein Sidedness

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Hydrophilic domains protrude from both

Most integral membrane proteins function in the following

© 2013 John Wiley & Sons, Inc. All rights reserved.

Driven by van der Waals forces between amino

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

A cytoplsmic proten Lipid-anchored proteins

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Distribution of Integral Proteins: Freeze-Fracture Analysis

© 2013 John Wiley & Sons, Inc. All rights reserved.

• These segments typically

© 2013 John Wiley & Sons, Inc. All rights reserved.

– Fluidity: measure of ease of flow

Lipids exist in 2 states

© 2013 John Wiley & Sons, Inc. All rights reserved.

• Liquid-crystal membranes predominate

Structure of the lipid bilayer depends on the temperature:

• The Importance of Membrane Fluidity

• Maintaining Membrane Fluidity

• The shorter the fatty acyl chain of

Prepared By: Dr. AbdulAmeer Al-Laith

Cholesterol – affects membrane physical state; interacts with

A. Membrane fluidity provides a perfect compromise

© 2013 John Wiley & Sons, Inc. All rights reserved.

© 2013 John Wiley & Sons, Inc. All rights reserved.

D. Many of the most basic cell processes (cell

• Lipid bilayer can exist in a relatively

Cell fusion to reveal mobility of membrane proteins: