TDC2h

Channels & Pumps

November, 2020
TDC: Interrelationship of lecture blocks

Receptors

Structural proteins and

Block 2 extracellular matrix
DNA RNA Proteins Nuclear enzymes and
Block 1 transcription factors
Cytosolic enzymes, motors,
pumps, channels

IMPORT/EXPORT
 Outline
• Channels - the K+ channel
• Ions and membrane potential
• Physiological Role of K+ channel
• Structural basis for ion selectivity
• Structural basis for rapid transport

• Pumps – Major Facilitator Superfamily

• Transport with an ion
• Accumulation against a concentration
Nature of Cytoplasm

300 mg/ml protein

300 mg.ml
Filaments
-1

300 g.l-1
Crowded!
Problem - osmosis

+
+ +
+ + -
+ +
- -
- -
- - +
-
The solution-pump out Na+
+ + +ve
+

+
+ +
+ + -
+ -ve
+
- - -
- - + -
-
K+ acts as a counter ion

Na + + +ve
Na + Na

K + +
+ K+ +
+ + -
+ + -ve
- -
- -
- - +
-
K+ /Na+ at equilibrium

K + Na+
V=-60mV

K+ Na+
 Summary - Role K+ channel
Na+ pumped out so cell does not
burst ( proteins -many conterions)

K+
Na+ As Na+ increased outside, potential
V=-60mV works against its efflux. 2 K+ exchanged
K+ Na+
inwards for 3 Na+ to prevent this

K+ In turn K+ leaks out via K+ channels so

that [K+] inside reaches steady state
K+
Common architecture of
channels
Ion selectivity of K+ channel
 K+ channel
• Very selective ( exclude smaller!)

• Very Fast (as if membrane was not

there)
Filtering too large and too
small
Structure prokaryote K +

channel
Cross section through pore
Basis for selectivity
=K+ =Na+
Basis for speed
TDC: Interrelationship of lecture blocks
 Major Facilitator Superfamily
• Transport small
molecules
• With an ion – Na+ or
H+
• In all phyla
• 160 out of 4000
genes in E.coli
 Lactose Permease, LacY
• Best characterised
since mutants can be
more easily made in
E.coli
• Biochemical results
show an ordered
mechanism.
 LacY, Structure
• Inner facing
conformation
• Lactose in solvent
cavity facing
cytoplasm
• Cysteine crosslinking
suggest rocking
motion to face
outwards
 Mechanism
• Mutants show
residues involved in: Binding Lactose
• Binding Lactose H+ and Lactose

• Transporting H+
• Coupling H+ to lactose
transport

H+ only
Conformations biased by H+
Outward Conformation
Inwards Conformation
 Lactose permease
H+

E
+ NH 2
O - O
NH 2
R
O
-

E
cytoplasm
O
 Lactose permease

E
+ NH 2
O OH
NH 2
R
O
-

E
cytoplasm
O
 Lactose permease
S

E
+ NH 2
O OH
NH 2
R
O
-

E
cytoplasm
O
 Lactose permease

O
- E
NH2 S
R O
+

H +

NH2 O
-

E
cytoplasm
O
 Lactose permease

O
NH2 - E
R S
O
+

NH2
HO
E
O
cytoplasm
 Lactose permease

O
NH2 - E
R S
O
+

NH2
HO
E
O
cytoplasm
 Lactose permease

O
NH2 - E
R O
+

NH2
H+ O
- E
O
cytoplasm
 Lactose permease
H+

-
E
+ NH 2
O - O
NH 2
R
O
-

E
cytoplasm
O
 Lactose permease

E
+ NH 2
O OH
NH 2
R
O
-

E
cytoplasm
O
Mechanism (1)- Protonation at
extracellular side
Mechanism (2)- Sugar binding at
extracellular side
Mechanism (3)-Preinversion
Mechanism (4)-Inversion
Mechanism (5)-Sugar Leaves
Mechanism (6)-Hydrogen ion
Leaves
 Mechanism- Overall
periplasm

cytoplasm
1 2 3 4

8 7 6 5
Pushkar Y. Pendse,  Bernard R. Brooks, and Jeffery B. Klauda (2010) J Mol Biol.  404. 506–521.