Sect2 Lec2 1ppt

Bio130 Part 2, Week 2:
Membrane Transport of
Small Molecules
Reading: Alberts et al. 5th edition
Chapter 11, pages 651-671
1
Outline
Permeability of lipid bilayer
Passive and active transport
Classes of Membrane Transport Proteins:

Uniporters
Symporters and antiporters
ATPases
Channel Proteins
2
Lipid bilayer is permeable to:
• Movement via simple diffusion through the lipid bilayer
– 1) High concentration to low concentration

 i.e. down the concentration gradient
– 2) More hydrophobic or non-polar the molecule

 faster the diffusion across the lipid bilayer
3
Lipid bilayer is impermeable to:
 These require membrane proteins for transport
4
Membrane Transport Proteins
• multipass transmembrane proteins
– protein-lined path across cell membrane
 transport polar and charged molecule
– ions, sugars, amino acids, nucleotides, various cell
metabolites
• different cell membranes
– different transport proteins
• each transport protein is selective
 transports a specific class of molecule
could just transport ions, +ve ions, Ca2+ ions
-different levels of specificity as to how, what, and when
5
Passive and active transport
high to low
concentration
low to high - reverse
needs energy
Transport proteins: all of the above proteins
TransportER proteins: not channel proteins
6
Resting Membrane Potential
outside + + + + + + + + + + + + + + +
Plasma membrane
_______________
inside
Positive molecules, uncharged, then negative charges has the highest to lowest motive force
across this membrane
7
Concentration gradient and
membrane potential
Work against
Additive
each other
works with membrane potential works against each other
concentration gradient + membrane potential (electrical gradient) = electrochemical gradient
*consider concentration AND charge when dealing in transport
8
Passive and active transport for
molecules with a net charge
electrochemical
gradient
9
Have a look at this animation
on the Bio130 Website
Carrier = Transporter
10
Types of Active Transport
• Active transport is against
electrochemical gradient
 needs energy
1) Coupled Transporters
 one molecule down gradient
 two molecules against gradient
2) ATP-driven pumps (“Pumps” or “ATPases”)

 ATP hydrolysis; moves molecules against gradient
3) Light-driven pumps (bacteria)

 light energy; moves molecules against gradient
11
Overview of Transport Proteins
Transporter proteins are carrier proteins!!
• A) Transporter Proteins
– a) Passive transport by transporter proteins
– I. Uniporter
– b) Active transport by transporter proteins
• 1) coupled transporters
– I. Symporter
– II. Antiporter
• 2) ATP-driven pumps (transport ATPases)
– I. P-type ATPase
– II. F-type and V-type ATPase
– III. ABC Transporters
• B) Channel Proteins (passive transport)
12
A) Transporter Proteins
• bind specific solute
 conformational change
• transport solute across membrane
13
– a) Passive transport by transporter proteins GLUT
– I. Uniporter Uniporter
– I. Symporter Na+/glucose symporter
– II. Antiporter Na+/H+ antiporter
14
Uniporters
Uniporter
– one molecule
 passively transport down electro-
chemical gradient Electrochemical
gradient
 direction of transport is reversible
if the gradient is reversed, it'll go in reverse
– eg. GLUT Uniporters
15
How a Uniporter Works
• Passive transport by a transporter
= facilitated diffusion
Animation:
http://highered.mcgraw-
hill.com/sites/0072495855/student_view0/chapter2/animation__how
_facilitated_diffusion_works.html
16
GLUT Uniporter
Blood vessel
Glucose
Electrochemical
gradient
cytosol
 transports glucose down the EC gradient
 works in both directions (glucose in or out of the cell)
17
– a) Passive transport by transporter proteins GLUT
– I. Uniporter Uniporter
– II. Antiporter Na+/H+ exchanger
18
18
Symporters & Antiporters
Symporters: two molecules
 moved in the same direction
Antiporters: two molecules
 moved in the opposite direction
19
THEY USE ENERGY, JUST NOT ATP!!!
Symporters & Antiporters cont’d
electro- electro- electro- electro-

chemical chemical chemical chemical
gradient gradient gradient gradient
low to high high to low
Modified from
uses energy from
Free energy from co-transported ion moving down THIS molecule
electrochemical gradient used to transport the 2nd molecule:

 against its EC gradient
 secondary active transport
20
Example: Na+/glucose symporter
Na+ down electrochemical gradient

 provides the energy to Where does antiport get energy from?
antiporter get the energy?
How does
 move glucose against the concentration gradient

Cooperative binding of Na+ and glucose leads to a
 conformational change in the protein
21 Co-operative binding - when one binds, the other is much more able to bind
Regulating Cytosolic pH
Most proteins require a specific pH
Cytosol: neutral ~7.2
Lysosomes: Acidic ~5, with excess H+
Excess H+
 leaks into the cell
 produced by acid forming reactions
Na+ driven antiporters maintain cytosolic pH.

22
Na+/H+ exchanger (Antiporter)
Na+
Uses the free energy stored in
extracellular space the Na+ electrochemical
gradient to move H+ out of the
cell.
Na+ electro- H+ electro-
chemical chemical
gradient gradient Responds to cytosolic pH
•pH drops (acidifies)
cytosol
•Transporter activity?
H+  Increases
23
Glucose Na+
Na+
extracellular space
cytosol
eventually sodium
will be equal... H+
Na+/glucose symporter Na+/H+ exchanger
*Specific for these molecules, ONLY.
These two transporters use the energy stored in the Na+

electrochemical gradient to move other molecules against
their electrochemical gradients
•Continued action would equalise the Na+ gradient
•How is the Na+ electrochemical gradient maintained?
 A transport ATPase, the Na+/K+ pump

24
– I. Uniporter GLUT
Uniporter
– I. P-type ATPase Na+/K+ pump
25
P-type ATPases
Animal plasma membrane Na+/K+ pump
• P-type transport ATPases extracellular space
 phosphoryates itself during

pumping cycle
• Na+ and K+
– moved against
electrochemical gradients
(see Table 11-1)
- uses actual ATP this time
Na+ gradient
 transport of nutrients into cells (e.g. Glucose)
 maintenance of pH, cell volume
26
How the Na+/K+ pump works
• http://highered.mcgraw-
hill.com/sites/0072495855/student_view0/ch
apter2/animation__how_the_sodium_potassi
um_pump_works.html
27
phosphorylating itself
Figure 11-15 cleaved into ADP

p. 662
Extracellular space
cytosol
Figure 11-15 Molecular Biology of the Cell (© Garland Science 2008)

• A) Carrier Proteins
– a) Passive transport
Uniporter
– b) Active transport
• 1) coupled carriers
– I. Symporter
Na+/glucose symporter
– II. Antiporter
29
Transport proteins work together to transfer
glucose from the intestine to the blood stream
inside; *apical side faces inside of lumen

food here
- prevents any movement between cells
needs energy, against

gradient
glucose uniporter;
no need for energy
basal-lateral
domain
outside; rest
of the body
30
Transcellular Transport of Glucose Requires the Asymmetric
Distribution of Membrane Proteins
• Intestinal epithelial cells

– transport proteins  restricted by tight junctions
• apical membrane  Na+/glucose symporter
• basolateral plasma membrane  GLUT2 uniporter and Na+/K+
pump
31
Uniporter
– I. P-type ATPase Na+/K+ pump help establish a gradient harnessed by
coupled transporters
– II. F-type and V-type ATPase important for resting membrane potential
– III. ABC Transporters osmotic balance

32
ATP-driven pumps (transport ATPases)
I II III
33
2) ATP-driven pumps (transport ATPases)
II. F-type and V-type ATPase
F-Type (e.g. ATP synthase):
- use the H+ gradient to drive the
- synthesis of ATP
Structurally related
- mitochondria, chloroplasts, bacteria
Opposite in terms of
modes of action
V-Type (e.g. H+ pump):
- use ATP
- pump H+ into organelles, acidify the lumen
- lysosome, plant vacuole
34
ATP synthase H+ Pump
F-TYPE PUMP V-TYPE PUMP
+ + + +
+ + + + + +
+ + + +
In mitochondria In lysosome - need pH of ~5
wants more H+
Inner membrane space Lysosome lumen
matrix cytosol
+ +
Uses the H+ electro- Uses ATP to pump H+

chemical gradient to against the electro-
produce ATP chemical gradient
(F-type) (V-type)
35
ATP Synthase Structure and
Mechanism
• http://www.dnatube.com/video/104/ATP-
synthase-structure-and-mechanism
36
ABC = ATP Binding Cassette:
 each member of this family contains two conserved ATPase domains
 pump small molecules across membranes
e.g. MDR (multidrug resistance protein)
 pumps small molecules out of cells
 overexpressed in cancer cells, drugs are pumped out.
37
Figure 11-17b Molecular Biology of the Cell (© Garland Science 2008)
Uniporter
• 1) coupled transporters Na+/glucose symporter
– I. Symporter Na+/H+ antiporter
– II. Antiporter
– II. F-type ATPase ATP Synthase
V-type ATPase H+ pump (some organelles)
– III. ABC Transporters MDR
• B) Channel Proteins (passive transport) 38

38
NOT TRANSPORT*ER* PROTEINS
B) Channel Proteins
• hydrophillic pore
across membrane
• most channel proteins are selective
– e.g. Ion channels
varying degrees of selectivness
• passive transport
 weak interactions with the solute
 faster transport by channel than by transporters
 several molecules pass through when open
39
Ion channels
Found in animals, plants, microorganisms
Two types:
1) Non-gated – always open
e.g. K+ leak channels. Major role in generation of resting
membrane potential in animal cells
2) Gated – chemical or electrical signal required

for channel opening
40
Non-Gated Ion Channels
1) Non-Gated Ion Channels
e.g. K+ Leak Channels
 always open
 potassium moves out of the cell down EC gradient
– plasma membrane of animal cells

• generates membrane potential across plasma
membranes (animals)
41
41
Membrane Potential
membrane potential =
 sides
difference in electrical charge on two
of the membrane
*Note: it's not the phospholipids or the
membrane itself that's charged - there are
just ions around the site creating the charge
Importance (e.g.):
used by symporters and antiporters to carry out secondary
active transport (animals and plants)
action potentials in nerve cells (animals)
42
42
Generation of Membrane Potential:
Plasma Membrane of Animal Cells
i) K+ Leak channel
 major role in membrane potenial
• outward flow of K+
ii) Na+/K+ pump

• Na+ gradient with low cytosolic [Na+]
• K+ gradient with high cytosolic [K+]
• Electrogenic
– 3 (+) ions pumped out Na+
– 2 (+) ions pumped into cell K+
 Net: 1 (+) ion pumped out
 ~10% of membrane potential
43
43
Membrane Potential:
Plasma Membrane of Animal Cells – cont’d
outside inside
Principles:
1) ions in solution - present in pairs
 each ion needs a counterion; +ve and -ve
• Outside: Na+; Cl-
• Cytosol: K+; Cl- & cell’s fixed anions (proteins, RNA,
other macromolecules)
2) ions diffuse from [high] to [low]
• K+ out → K+ leak channels
Net result: ++++ ++
 more (+) on ouside (Na+, leaked K+)

--- - --
 more (-) on inside (Cl- and fixed anions)
• forms membrane potential
44
44
Generation of the membrane
potential in animal cells
+_ +_ +_ +_ +_ _+ +_ +_
3Na+ _ + Na+/K+ pump
_
2K + +
_ _ +
+
[Na+] + _
_ [K ]
+
_
_ +
+ +
[Cl-] _
-,anions]
_
+
_ [Cl _
+
+
_ + K+ leak channel
K+ _ +
+ _ _ _ _ _ _ _ _ +
+ + + + + + +
Net result:
 more (+) on outside, Na+ and leaked K +
more (-) on inside, Cl- and fixed anions
45
forms membrane potential
Membrane Potential
http://www.dnatube.com/video/5041/The-Resting-
Membrane-Potential
46
Membrane Potential:
Plasma Membrane of Animal Cells – cont’d
Resting Membrane Potential

• high [K+] in cytosol
– outward flow of K+ → K+ leak
channels
• balanced by
 repulsion of the excess (+) on outside
 attraction to excess (-) on inside
• equilibrium = resting membrane potential
• animal cells: ++++ ++
vary from -20 mV to -120 mV

--- - --
47
47
Gated Ion Channels - Types
i) voltage-gated ion channel ii) mechanically-gated ion channel
 changes in voltage across membrane
activates or inactivates them
 mechanical stress; physically moved
• opens if plasma membrane
stretched
48
Gated Ion Channels - Types
iii) ligand-gated ion channel iv) ligand-gated ion channel
(extracellular ligand) (intracellular ligand)
e.g. extracellular ligand e.g. intracellular ligand
 neurotransmitters  ion, nucleotide
outside
inside
inside
49
An Example of a Ligand-gated Ion Channel
Converts chemical signal into

 an electrical signal
Open in response to
neurotransmitter
- ions flow through the channel
- local changes in membrane potential
 opens voltage gated channels
50
Figure 11-35a Molecular Biology of the Cell (© Garland Science 2008)
Next Week: Intracellular
compartments and protein sorting
Tutorial: Membrane Transport of

Small Molecules
51

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Bio130 Part 2, Week 2:

Permeability of lipid bilayer

Passive and active transport

Classes of Membrane Transport Proteins:

• Movement via simple diffusion through the lipid bilayer

– 1) High concentration to low concentration

– 2) More hydrophobic or non-polar the molecule

 These require membrane proteins for transport

low to high - reverse

works with membrane potential works against each other

concentration gradient + membrane potential (electrical gradient) = electrochemical gradient

*consider concentration AND charge when dealing in transport

 two molecules against gradient

2) ATP-driven pumps (“Pumps” or “ATPases”)

3) Light-driven pumps (bacteria)

 direction of transport is reversible

if the gradient is reversed, it'll go in reverse

– eg. GLUT Uniporters

 transports glucose down the EC gradient

 works in both directions (glucose in or out of the cell)

Symporters & Antiporters cont’d

electro- electro- electro- electro-

low to high high to low

electrochemical gradient used to transport the 2nd molecule:

 secondary active transport

Na+ down electrochemical gradient

 move glucose against the concentration gradient

Na+ driven antiporters maintain cytosolic pH.

These two transporters use the energy stored in the Na+

 A transport ATPase, the Na+/K+ pump

• P-type transport ATPases extracellular space

 phosphoryates itself during

Figure 11-15 cleaved into ADP

Figure 11-15 Molecular Biology of the Cell (© Garland Science 2008)

inside; *apical side faces inside of lumen

- prevents any movement between cells

needs energy, against

• Intestinal epithelial cells

• B) Channel Proteins (passive transport)

Uses the H+ electro- Uses ATP to pump H+

• B) Channel Proteins (passive transport) 38

2) Gated – chemical or electrical signal required

 potassium moves out of the cell down EC gradient

– plasma membrane of animal cells

ii) Na+/K+ pump

Net result: ++++ ++

 more (+) on ouside (Na+, leaked K+)

Resting Membrane Potential

vary from -20 mV to -120 mV

Converts chemical signal into

 opens voltage gated channels

Tutorial: Membrane Transport of

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