Cell Biology Tutorial 2-The Plasma

Membrane
Draw a diagram (with dimensions in nm) of the
fluid-mosaic model of the plasma membrane.
Label your diagram to include:

a) phospholipid d) integral (intrinsic) protein
b) sterols e) peripheral (extrinsic)
protein
c) glycolipid f) glycoprotein


Width

= 6-10nm

Glycolipid
(showing
sugar
residues)
Peripheral protein bound to
integral protein by polar forces
Phospholipid with internal hydrophobic
acyl chains; hydrophilic functional
groups (ie polar head groups)
orientated outwards
Integral
protein
Glycoprotein
(showing
sugar residues)
Width

= 6-10nm
Sterol (eg cholesterol*) with
appropriate
charge orientation
*Cholesterol only found in
eukaryotic cell membranes
Summarise the evidence for its
asymmetrical structure.

The membrane is not homogenous:
different parts of the membrane have
different functions
Evidence for asymmetrical
structure includes:

Cell markers with high affinity for sugar residues can be seen
bound to the outer surface of permeabilised cells using
microscopes.
Freeze-fracture studies.
Enzyme systems / ion pumps in membranes are asymmetrically
orientated, eg Na-K pump only inhibited by ouabain from the
outside; ATP must be supplied intracellularly
Cell surface staining-Antibody
binds to the outer surface only
Freeze Fracture splits the lipid
bilayer
Freeze fracture Image
Na
+
/K
+
pump
3 Na
+
(OUTSIDE CELL)
(INSIDE CELL)
2 K
+
Ouabain inhibits at K+
binding site
ATP
(energy to
drive
pump)
ADP + Pi
Vanadate inhibits ATPase
Link Barrier function of cell membrane
How do
lipids,
proteins
carbohydrates
contribute to the link-barrier function of
the membrane?
Link Barrier function of cell membrane
Lipid:
permeability barrier for hydrophilic molecules;
solvent for integral proteins;
functional roles for glycolipids, eg insulator in myelin
sheaths.
Protein:
transport, communication, energy transduction.
Carbohydrate:
intercellular recognition (eg. blood group factors); may help
maintain protein (internal)
orientation of glycoproteins.

Membrane composition relates to
function

How does the composition of mammalian
and bacterial cell membranes differ?
Membrane composition relates to
function
Bacterial plasma membrane have:
a higher protein content (~75%) than most
eukaryotic plasma membranes (~50%)
a different lipid spectrum more
phosphatidyl ethanolamine and lack
cholesterol

Circle the correct term(s) in the
following:
a) the life of most membrane components is
roughly 50 secs/50 hours/50 days.
b) transverse diffusion of membrane
components is/is not rapid.
c) lateral diffusion of membrane components
is/is not rapid.
d)membrane fluidity is controlled by its
lipid/protein/carbohydrate composition.


Circle the correct term(s) in the following:

e) membrane components are held together by
covalent/non-covalent forces.
f) plasma membranes have a high/low
permeability for most charged molecules.
g)plasma membranes have a high/low
permeability for water
h)plasma membranes have a high/low
permeability for respiratory gases.


We will now look more
carefully at the answers to
questions a to e
Circle the correct term(s) in the following:
a) the life of most membrane components is roughly 50
secs/50 hours/50 days.

Example: Turnover of membrane components (liver). The Half life is
the time for 50% of the component to be removed (and replaced)







Structure Half life (T
1/2
,hours)
phospholipid 50
Proteins 50
Nuclear proteins 120
cholesterol 60
b) transverse diffusion of membrane components is not rapid
Very slow
Transverse diffusion of phospholipid (flip flop)
takes place about once a month whereas lateral
movement ocurs 10
7
times a second.
Hydrophobic
interior
c) lateral diffusion of membrane components is rapid.



Rapid
Lateral diffusion 2 m s
-1
;
( 2 m approximate length of bacterium)
Proteins vary in their lateral mobility depends
upon cytoskeletal attachment
Membranes are about as fluid as lightweight machine oil.

A given phospholipid in a bacterial plasma membrane
may travel from one end of the bacterium to the other in
approx 1 second
d) membrane fluidity is controlled by its lipid/protein/carbohydrate
composition.
Membrane fluidity controlled by fatty acid composition and
cholesterol content
Prokaryotes regulate fluidity by varying the number of double
bonds and length of fatty acyl chains; double bonds put akink
into the chains
Eukaryotes regulate fluidity by introducing cholesterol.; bulky
molecule separate the fatty acyl chains.
e) membrane components are held together by covalent/non-
covalent forces.

Hydrophobic attraction: non-polar molecules tend to cluster
together in water to minimise the number of exposed
hydrocarbon chains
Van der Waals forces: Attractive forces which come into play when
any two atoms are 3-5 apart. Becomes significant when
numerous atoms in one molecule can approach many atoms in
another; (steric complementarity)


f) plasma membranes have a high/ low
permeability for most charged molecules.
g) plasma membranes have a high/ low
permeability for water (water passes through
channels integral proteins called
aquaporins).
h) plasma membranes have a high /low
permeability for respiratory gases.
5) Distinguish between
a) passive diffusion
b) facilitated diffusion,
c) active transport,
d) bulk transport of
metabolites across
membranes.

a) passive diffusion
molecules or ions go down
concentration gradient
no carriers or special channels
required
no ATP required.

b) facilitated diffusion
facilitated transport = facilitated diffusion
Molecules or ions go down a concentration
gradient
but via pores or channels
no ATP required.

c) active transport
molecules or ions go up or against a
concentration gradient
carriers required
ATP required (by ATPases),
d) bulk transport of metabolites across
membranes
movement of materials in bulk under
pressure as occurs across capillary
membranes
6) Outline the distribution of the major
anions and cations across the plasma
membrane and state the local charge
separation which always exists across
the plasma membrane.

Comparison of the composition of intracellular fluid (ICF)
and extracellular fluid (ECF).


Cations Anions Cations Anions
K
+
155 Cl
-
8 K
+
5 Cl
-
103
Na
+
12 HCO
3
-
8 Na
+
142 HCO
3
-
27
Mg
+
15 Protein
-
60 Mg
+
3 Protein
-
16
Ca
2+
2 PO
4
2-
90 Ca
2+
5 PO
4
2-
2
Others 7
Total 184 Total 184 Total 155 Total 155
Ions in ICF (mmol dm
-3
)
Ions in ECF (mmol dm
-3
)
Differences in ion concentration
across membranes occurs because


differential membrane permeability to
ions

activities of transport processes/ion
pumps.

-ve Proteins /phosphates
(cannot cross membrane)
[K
+
]
[K
+
]
[Cl
-
]
[Cl
-
]
3 Na
+
2 K
+
Pump
Equilibrium membrane
potential
(-70mV in nerve cells)