Biomolecular

Membrane
Dra Sri Mustaina, MKes

Functions of Membrane
1.

Compartmentalization

2. Selectively permeable membrane barrier

3. Transport phenomena
4. communication and compartment attachment
5. Cell identity and antigenicity
6. Conductivity

The Fluid Mosaic Model

Sheet-like non-covalent assemblies
Regulates transport of small and macromolecules
Controls flow of information between cells
Specific receptors for external stimuli
Generation of signals
Cell adhesion for tissue formation.
Energy conversion processes- a battery.

- Small platform, composed

of sphingolipids, chol &
prot
- fluid, more ordered &
tightly packed.

- function : regulate membrane functions

- Prot : GPI anchored, tyrosin kinase, G
protein fragments, eNOS
- Disease : alzheimer, muscular disthrophi,
astme, allergic respons, etc.

Raft clustering.

Tr
a
n
s
p
or
t
lI
nt
as
m
e
m
br
a
n

Overview of transport mechanisms

Plasma membrane is a semi-permeable
membrane
Small hydrophobic
molecules: O2, CO2, N2,
benzene
Small
uncharged polar
molecules: H2O, ethanol,
Lipid
glycerol
bilayer
Larger uncharged polar
molecules: glucose,
amino acid, nucleotides
Ions: H+, Na+, HCO3- ,
K+, Ca+, Mg2+, Cl- , etc.

DIFFUSION (hydrophobic/
lipophilic molecules only):
a slow process.

FACILITATED
DIFFUSION, uniport (e.g.
glucose transporters)

CO-TRANSPORT:
symport (same direction) or
antiport (opposite directions

CHANNELS, e.g. sodium

channel, water channel
(aquaporin)
ATP

ACTIVE TRANSPORTER,
ADP + Pi against the concentration
gradient

Diffusion :

kinetic energy

Diffusion rate:

temperature
distance
concentration gradien
area
Determined by : molecular size + attribute
solvent characteristic

Simple diffusion :
- oxygen
- carbon dioxide
- water
- uncharged
molecules
- lipid soluble

OSMOSIS

diffusion of water molecules through a

selectively semipermeable membrane

from hypoosmotic sol to hyperosmotic sol

continues until :
equilibrium reached
cell destroyed
stopped by opposing force

SOLVENT FLOW.
Movement of water occurs by one of two
processes.
1. Osmosis
2. Filtration
(Example: urine formation in kidney)
3. Specific Channel

Aquaporin
water channel proteins called Aquaporins (AQP1 & AQP2)

Passage of water molecules through the aquaporin AQP1. Because of the positive charge
at the center of the channel, positively charged ions such as H3O+, are deflected. This
prevents proton leakage through the channel.

Peter Agres experiment with cells containing or lacking aquaporin. Aquaporin is

necessary for making the 'cell' absorb water and swell.

Facilitated diffusion
Molecules with low permeability coefficients can go

through the membrane faster than normal diffusion

process.
Ex : uptake of glucose into erythrocyte.
It is rapidly moved across the membrane down the
concentration gradient with the help of a membrane
protein called permease.
The velocity of transport is saturable
Permease is a multipass transmembrane protein
to facilitate the diffusion of specific molecules across
biological membrane.

Transport
velocity

Glucose
transporter
(permease)
ATP

ADP

Rate of Glucose Uptake

Facilitated Diffusion: with molecule specific

permease

Saturable
Vmax
Facilitated
diffusion
Half Vmax
Simple diffusion

Km
External concentration of glucose,
mM

Transporters must have a specific binding site for the

solute, e.g. glucose. Once they get in the cells, they are
usually phosphorylated to go into metabolic pathway and
hence the outer concentration of glucose is higher than
inside.

Facilitated

Diffusion of Ions

The transmembrane channels that permit facilitated diffusion can

be opened or closed. They are said to be "gated
1. Ligand-gated ion channels.
open or close in response to binding a small signaling molecule or
"ligand". (extracellular ligands and intracellular ligands) The
ligand is not the substance that is transported when the channel
opens.
2. Mechanically-gated ion channels
Examples: Mechanical deformation of the cells of stretch receptors
opens ion channels leading to the creation of nerve impulses.
3. Voltage-gated ion channels
In so-called "excitable" cells like neurons and muscle cells, some
channels open or close in response to changes in the charge
(measured in volts) across the plasma membrane.

Channels
Voltage gated
channels

NH2

COOH

NH2

Sodium channel consists of

FOUR transmembrane domain,
each has SIX transmembrane
helices, the forth helice is
believed to be the voltage

COOH

Potassium channel
has ONE molecule of
only SIX
transmembrane
helices

Active transport

Concepts of active transport

Use ATP hydrolysis directly or indirectly (secondary

active transport) to move molecules across the

membrane against the concentration gradient.
Active transporters are membrane proteins
specifically bind and move the molecules across the
membrane to a unique direction using ATP
hydrolysis as an energy source.
Mainly for ions and chemicals, creating a chemical
gradient using ATP energy.
The gradients created maintain many life processes.

TRANSPORTER
specific
conformation changes
slower than Channel protein
3 tipe : - passive transporter
- active transporter
- group transporter
- Mechanism

: - uniporter
- antiporter
- symporter

Tipe Transporter
UNIPORTER
1 molekul, searah gradien konsentrasi
c/ : glukosa transporter

SYMPORTER
tanpa ATP, energi dari gradien
melibatkan 2 jenis molekul
sama dengan kerja pompa
arah : sama, c/ : Na+ dan glukosa, di lumen usus-jaringan

ANTIPORTER
arah : berlawanan c/: Na+ dan Ca+, di otot jantung

Indirect active transport

P-class pump
Perubahan konformasi protein yang disebabkan
karena proses fosforilasi oleh ATP
F-class / V-class
Proton transporting protein (H+).
V-class : mempertahankan pH, pada lisosom, dan
vesikula lain
F-class : di mitokondria, melawan gradien elektrokimia

Three types of ATPases: P, V & F.

1997 Nobel prize of Physiology and Medicine went to Paul
Boyer and John Walker for their work on ATP synthase (F type),
and Jens Skou for his work on Na-K-ATPase, which uses onethird of the ATP made by ATP synthase.
V-type ATPase

F1

P-type ATPase

F-type ATPase

complex

V1
comple
x

F0
complex

V0
complex

Proton
pump

ATP
synthase

Classified according to their protein sequence homology and structures.

P-type: Na+-K+-ATPase, Ca2+ and H+ pump, P means they

have phosphorylation and they all sensitive to inhibition.

V-type: inner membrane ATPase to regulate H+ and adjust

proton gradients, v means vacuole type for acidification of

lysosomes, endosomes, golgi, and secretory vesicles.

F-type: ATP synthase to generate ATP energy from moving

the proton across; F means energy coupling factor. There are

F1 and F0 subcomplexes: F1 generates ATP, F0 lets H+ go
through the membrane.

ABC transporters: ATP-binding cassette protein for active

transport of hydrophobic chemicals and Cl-.

P-type ATPase: Ca2+ATPase

Ca2+

Ca2+ binding
site
Aspartate
ATP binding
site

2
NH

COO
H

ATP
ADP

Other divalent ion transporters have similar structure

with this Ca2+-ATPase and the subunit of Na+-K+ATPase.

ABC (ATP-Binding Cassette) transporters: 6

or 12 trans-membrane helices with 2 ATP binding
sites, drug or ligand- binding sites yet to be clearly
identified.
Chloride channel: the cystic fibrosis
P-glycoprotein
transmembrane conductance regulator,
CFTR, has an extra R domain

CO
OH

CO
OH

NH2

NH2

Oligosaccharid
e chains

R
domain

ATP binding domains

ATP binding domains

Vesicle-mediated transport

Vesicle-mediated transport

Vesicular Transport : Receptor Mediated Endocytosis

-

receptor-mediated endocytosis: requires

specific receptor/ligand pair
small pit contains ligand receptors exposed
to extracellular space and adaptin and
clathrin on the intracellular side (skeleton
for vesicle)
adaptin links receptor (intracellular side) and
clathrin to form a coated pit
clathrin forms polyhedral cage giving the
"coat" of the vesicle
pinching off of vesicle into cell is requires
dynamin,a GTPase
clathrin-coated vesicle is "de-coated" of
clathrin and the proteins involved in
formation are recycled while the vesicle is
further processed
phagocytosis:ingestion of particles (bacteria,
dust, carbon particles)

Junction
A. CELL TO CELL JUNCTION.
1. OCCLUDING JUNCTION / TIGHT JUNCTION
2. ADHERING JUNCTION : actin filament = ZONULLA
intermediate filament = MACULA
B. CELL TO CELL COMMUNICATION : gap junction
C. CELL MATRIX / ANCHORING JUNCTION
Not only hold cells together but provide tissues with structural cohesion. These
junctions are most abundant in tissues that are subject to constant mechanical
stress such as skin and heart.
1. FOCAL ADHESION / DESMOSOME
plasma membrane - plasma membrane of adjacent cells.
2. HEMI DESMOSOME
cytoskeleton and extracellular matrix components such as the basal
laminae that underlie epithelia

Occluding junction : tight junction

Adhering junction :
a. Zonula adherent
b. Macula adherent

Cell cell communication

Gap junction

Cell-matrix Adhessive junction : anchoring junction

Desmosome

hemidesmosome

Anion antiport in parietal cells of stomach

with H+- K+ -ATPase to produce stomach
acid.
Cl -channel

Anion antiport
-

Cl
HCO3

Cl

HCO3
Carbonic
anhydrase

CO2

Cl -

CO2
+ OH
-

Basolateral
membrane

ATP

K+

ADP + Pi

H 2O

K+
K+

H+-K+ATPase

Omeprazole
inhibits the
+
K proton
channel
pump.

Apical
membrane

Omeprazole and Cimetidine stop

stomach acid
H+-K+-ATPase is an electroneutral antiport. K+ is

removed by K+ channel and concurrently Cl- channel

removes Cl- to the same direction.
HCl is the overall transport product in the stomach
lumen.
Omeprazole inhibits the proton pump directly.
Cimetidine resembles histamine to block the binding
of histamine to its receptor thus inhibit the activation
of H+K+-ATPase by histamine receptor.

Glucose uptake from lumen to the capillaries

using glucose transporter (permease),
glucose-sodium symport and Na+-K+-ATPase.
Intestinal
lumen

Na+
Na+-Glucose
Symport

Glucose
transporter:
permease

Glucose gets into

the cell and
Capillarie
simultaneously
s
transported by
glucose
transporter into
the capillaries.

ATP

Na+
K+

Na+
ADP +
Pi

Glucose-Na+
symport is driven
by intracellular
Na+ levels using
Na+ K+ -ATPase.

K+
Na+-K+ATPase

Co-transport Systems and

their coupling
K+ Na+
Na+ -K+
ATPase

Na+

solute
Na+ -driven symport

ATP
K+ = 4 mM

K+

Na+ = 145 mM

ADP +
+
NaPi

solute

Na+

K+ = 140 mM

H+

Na+ = 12 mM

+
+
+
+

H+
ATPase

ATP

ADP +
Pi
H+

Primary and secondary active transporters work

coordinately in animal cells. They generate membrane
potential, generate proton gradient, maintain acidity,

Na+-K+-ATPase and Ca2+-ATPase

Ca-channel
Ca-ATPase

Ca-Na Antiport

Na +

Na +
Ouabain
inhibits
Na-KATPase.

Na-K ATPase

Adapted from
from Sperlakis
Sperlakis (ed).,
(ed)., 1998.
1998. Cell
Cell
Adapted
Physiology Source
Source Book.
Book. Academic
Academic Press.
Press.
Physiology

K+

Sarcoplasmi
c reticulum

Ouabains for treatments of angina

pectoris and myocardial infarction
Ouabain blocks Na + -K + -ATPase. By blocking

the Na+-K+-ATPase, Intracellular Na+ remains

high
Hence, the Na+- Ca2+ antiport cannot remove
Ca2+ ions out from the cardiac muscle cells.
Eventually, the Ca2+ ion level is restored to
maintain the contraction power of cardiac muscle.

MEMBRANE & DISEASES

Can be disrupted by :
- Change to protein
- component of the lipid bilayer : protein
lipid
composition
- component of cytoskeleton
-

Categories of true membrane based

disease :
1. Defects in cytoskeletal component
impair membrane function
2. Altered membrane lipid composition
disrupts membrane traficking.

Defects in cytoskeletal component and

membrane function :
Ex.
-Sickle cell anemia
Actin/spectrin lattice lock, RBC less drformable and obstruct
the microcirculation

- Duchene muscular dystrophy

Mutation in dystrophin gene disrupts the ability of protein product
to anchor cytoskeletal element to the surface membrane.
Structural support is loss, membrane becomes permeable, intracell
pressure

and cell explodes

DisrupTs of membrane trafficking

-Rab protein :
Lipid modification to carboxyl terminus.
-Hermansky-Pudlak, Griscelly sindrome
Lysosome-related organelle
-Niemann-Pick Disease type C (NPC)
LSD

Protein membran dysfunction

Cystic fibrosis
Defect pada mekanisme transport chlorida di epitel paru
(CFTR = Cystic Fibrosis Transmembrane Regulator)

Pathologic condition :
CFTR transport Cl- : epithelial airway lumen,
followed by Na+ and water : thick & dehydrated
mucous in the lungs
Good breeding ground for infection by bacteria
Trouble breathing

Intracellular drug delivery

- Uptake by cell : 1. endocytosis
~ 500 nm
clathrin coated pits
pH environment
2. macropinocytosis, phagocytosis
scavenger (m, neutrophils) APC
size : up to the size of cell
- Cross the plasma membrane
- Escape from endosome/lysosome

Intracellular drug delivery

(cont)

- Cross the plasma membrane

Direct entry to cytosol
Viral peptide transporters
- Escape from endosome/lysosome
cargo released from vesicle once taken inside the cell
routes : - viral peptides evolved for endosomal escape
- fusion with endosomal membranes
- disruption of endosomal compartments