Molecular Cell Biology

Prof. D. Karunagaran
Department of Biotechnology
Indian Institute of Technology Madras

Module 7
Cell Signaling Mechanisms
Lecture 11
Ion Channels








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Ion channel facilitates the diffusion of ions via membranes in living systems.
Ion channels operate based on membrane potential/specific
ligands/concentration gradient

Types of ion channels
Substrate gated channels (Nicotinic receptor)
Simple pores (GAP junctions)
Pumps (ATP-synthase, K
+
,Na
+
-ATPase)
Voltage-gated channels (K-channels)

Mechanism of ion channel opening
Change in conformation occurs at a discrete area in the channel and causes
channel opening.
The entire channel changes in conformation
Neurotransmitter or hormone binding causes the channel to open
Neurotransmitter or hormone binding to receptor causes secondary messenger
activation mainly a protein kinase that phosphorylates a channel and opens it

Mechanism of Gating
Conformational changes in channel proteins are responsible for opening and
closing of the pore
Energy to switch the channel protein from one conformational shape to another
comes from the gating source





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Na
+
channel
Conducts impulses in nervous system
When the membrane potential reverses channel opens
Inflow of Na
+
ions takes place when the resulting local depolarization of the
membrane, which in turn activates neighboring voltage dependent Na
+
channels.
This phenomenon is known as action potential
K
+
ions are propelled out leading to repolarization of the membrane
Na
+
channel contains several subunits including alpha-subunit which mediates
Na
+
transport
Contains six transmembrane helices and S6 helices of all the domains forms a
hydrophilic pore at the center
Six S4 helices function as voltage sensors. This contains several positively
charged residues and during membrane polarization inner negative charges hold
the helix in the membrane.
When this attraction is removed due to local depolarization S4 helices move
upwards like springs resulting in opening of central pore.
Ligand gated channels
Binding of neurotransmitter causes flow of ions








= Neurotransmitter
Ion Flow = Current
3
Acetylcholine Receptor









Pentameric protein subunits, with two binding sites for acetylcholine, which, when
bound, alter the receptor's configuration and cause an internal pore to open and
allow Na+ions to flow down their electrochemical gradient into the cell
the resting (closed) ion channel to acetylcholine (ACh) producesthe excited
(open) state. Longer exposure leads to desensitization and channel closure.








Acetylcholine
binding sites
ACh
Na
+
, Ca
2+

Continued
excitation
Desensitized
(gate closed)
Excited
(gate open)
Resting
(gate closed)
Outside
Inside
ACh


ACh
(or )

ACh

4
Four types of transport ATPases
F-type
Reversible, ATP-driven Proton Pumps
P-type
Cation transporter that is reversibly phosphorylated as part of the transport cycle,
Na+, K+, Ca++channel
P-type ATPase maintains potassium and sodium gradient
V-type
Works as a proton pump
Has key role in acidification of cellular compartments (including endosome)
Multi-drug transporter
Export numerous compounds in ATP-dependent manner
Neurotransmission
Neurotransmission is rapid and action Potential opens voltage gated Ca
2+
channels
Ca
2+
enters the terminal and initiates vesicular release of neurotransmitter
Axonal conduction
Depolarization of the axonal membrane potential results in an action potential
The increase in action potential is due to sodium current flowing through voltage-
activated sodium channels
As the membrane potential decreases potassium ion flows out, this opposes
further depolarization and initiates repolarization
Longitudinal spread of local depolarizing sodium currents results in progressive,
longitudinal activation of sodium channels and new sites of depolarization. The
rate of conduction is dependent on the number and synchrony of sodium channel
activation
Number and synchrony of sodium channel activation are membrane potential
dependent
As the resting membrane potential decrease, fewer sodium channels will be
activated by a depolarizing influence and conduction velocity slows
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Synaptic Acti vity
Storage and release of neurotransmitter
Small molecule neurotransmitters (e.g. acetylcholine, norepinephrine) are
synthesized at axonal terminals and stored in synaptic vesicles

Toxins and Ion channels
Puffer fish: Tetrodotoxin (TTX)- Clogs Na
+
permeable pore
Red Tide: Saxitoxin- Na
+
Channel-blocking toxin
Batrachotoxin (frog): Blocks inactivation -channels remain open
Veratridine (lilies): Inactivates channels
Aconitine (buttercups): Inactivates channels
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Patch-clamp Technique
Patch-clamp conductance measurements reveal activities of single channels
Ion channels are best studied by this method
This technique monitors flow of ions through channels



Study Questions
1. What are the types of ion channels?
2. Write a note on different domains of Na+channel
3. V type works as to
a) proton pump b) sodium pump c) potassium pump d) none of the above
4. Match the following
Aconitine pump
Acetycholine Voltage channel
K-channels Inactivates channels
ATPase Opens channels
5. ..is used to Monitor flow of ions through channels
Cell

Channel

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