Neurobiology 15-BIOL-540 Winter, 2012 Lecture 7

Bacterial K+ channel.

extracellular space
H5

The first K+ channel for which an X-ray crystal structure was solved is from bacteria (Streptomyces lividans).

+
1 2 3 4 5 6

+
N cytosol C

In this channel protein: There are only 2 transmembrane a-helices, corresponding to helices # 5 & 6 in the mammalian voltage-gated channel. An intervening segment includes the K+ channel consensus sequence.

Bacterial K+ channel.

extracellular space
H5

Selectivity: K+ channels are highly selective for K+, e.g., relative to Na+.

+
1 2 3 4 5 6

+
N cytosol C

The selectivity filter that determines which cation can pass through a channel is located at the narrowest part. Mutation studies showed that the H5 segment is essential for K+ selectivity. H5 includes a consensus sequence (Thr-Val-Gly-Tyr-Gly) found in all K+ channels, with only minor changes through evolution.

Left: side view, in plane of membrane. Right: view from the extracellular space.

KcsA K+ channel: two views

PDB 1J95

The K+ channel consensus sequence (in black) forms the selectivity filter at the narrowest part of the channel. K+ ions are bound within the selectivity filter (color pink). The gate is where the innermost helices of the 4 subunits meet at the narrow end of the "teepee" structure. This is the closed channel.

extracellular space
H5

Voltage sensing: In mammalian channels (+) residues in helix #4 are essential for voltage gating. In helix #4 every 3rd residue is Arg or Lys, & intervening residues are hydrophobic.

+
1 2 3 4 5 6

+
N cytosol
H H COO H3N+ C CH2 CH2 CH2 CH2
+ + NH

Decreased transmembrane potential causes helix #4 to change position, resulting in more of its (+) charges being accessible to the aqueous phase outside the cell. A small "gating current" is measurable, as (+) charges effectively move outward.

H3N+ C CH2 CH2 CH2 NH C NH2

COO

NH2

arginine

lysine

voltage-sensor paddle

K+ in selectivity filter

side view within membrane facing membrane Voltage-gated K+ Channel

PDB 1ORQ

Some helices of the voltage-sensing domain of the differ from the expected trans-membrane orientation.
The (+) charged voltage-sensing helix #4, & part of what was designated helix #3, form a "paddle" shape at the periphery of the channel.

A look at the proteins responsible for generating and maintaining membrane potential.

Na+ gradient Na+,Clgradient

K+gradient

Na+gradient

Na+gradient

Na+-K+ ATPase pump

Ca2+ pump

Cell interior [A+]=10mM. [A+]=100mM. [B+]=100mM. [B+]=10mM. [C--]=100mM. [C--]=10mM. [D--]=10mM. [D--]=100mM. (Other ions, too)

The cell is at a resting potential of -58mV. The membrane potential Vm will ______ after the following changes in membrane permeability to ions A, B, C, D:

PA is increased. PB is decreased.

Cell interior [A+]=10mM. [A+]=100mM. [B+]=100mM. [B+]=10mM. [C--]=100mM. [C--]=10mM. [D--]=10mM. [D--]=100mM. (Other ions, too)

The cell is at a resting potential of -58mV. The membrane potential Vm will ______ after the following changes in membrane permeability to ions A, B, C, D:

pA is increased. pB is decreased.

Sodium potassium pump