ION CHANNELS

I. Basic biophysical properties of ion channels

II. Structure and Function

III. Regulation by Subunits and Enzymes

IV. Physiological functions of ion channels

V. Ion Channels and Disease

VI. Ion Channels as Targets for Drugs

Book references: Important slides: !

Essential cell biology: ECB: 397-414 (4th edition)
Molecular Cell Biology: MCB Ch 7.7, Ch. 13
Molecular Biology of the cell MBC Ch11, Ch15
 Keywords
Ion channel gating
Ion channel selectivity
Charge transport function of ion channels
Ion transport function of ion channels
Voltage sensor, pore domain, selectivity filter
Auxiliary subunit
Phosphorylation regulation of Ca2+ channels
G-protein-regulated K+ channel
Chemical synapse
Glutamate receptor
NMDA receptor
Ionotropic acetylcholine receptor
Metabotropic acetylcholine receptor
Electric synapse
Long QT syndrome
KATP channel
Channelopathy
Local anesthetics as drugs, action on ion channels
Sedatives as drugs, action on ion channels
 REVISION
Ion channels are transmembrane transport proteins

closed open

ATP dependent pump Ion channel transporters

 REVISION
I. Basic biophysical properties
(review of biophysics course)

• Conduction
hydrophilic pores
through the membrane

Voltage-gated change in the membrane potential

Ligand-gated binding of extracellular ligand

• Gating i.c. signal-gated binding of second messengers

Stretch gated membrane stretch, mechanosensor
Backround channel always open, but modulated

Highly-selective K+, Na+, Ca2+, Cl-

• Selectivity Mildly selective nAch receptor, cation spec.

Non-selective Gap junction channel

II. Structure and Function
 MBC. Ch11 p672 MCB. Ch7 p282

Well defined functions are associated with various

structural parts of the channel proteins

Example: voltage-gated K+ channel

voltage sensor
detecting the changes of membrane
potential

pore region
forming the ion conduction pore

selectivity filter
responsible for high K+ selectivity of the
channel

four subunits make up a functional channel

 info
Voltage-gated ion channels
evolved from a common
ancestral gene.

Na+ and Ca2+ channels:

one continuous polypeptide
chain
K+ channels:
4 separate subunits

The properties of a K+ channel

constructed from two different
types of subunits will differ from
those of both homo-tetramers.
MCB. Ch7 p282
 info
Studying ion channels in expression systems
Purpose: the expression of the target channel in high numbers without the
presence of other channels for “cleaner” measurements

Xenopus laevis

removal
of oocytes
III. Regulation by Subunits and
Enzymes
! Auxiliary subunits may be associated with the pore-forming -
subunits to “fine tune” the operation of the channel

Example:
The skeletal muscle Ca2+ channel consists
of five subunits. The auxiliary subunits
modulate the magnitude and kinetic
properties of the Ca2+ current and confer
sensitivity to channel modulators.
pore-forming
increases the peak (principal) subunit
current, alters gating

Mutations that prevent the operation

of the  subunit result in a decrease
of the Ca2+ current
 Ion channels can be regulated by phosphorylation !
Phosphorylation is a widely used regulatory mechanism, in which
a phosphate (PO4) group is attached to a protein modifying its
function in a reversible manner. Kinases are enzymes that add the
phosphate group and phosphatases are enzymes that remove it.

dephosphorylated phosphorylated
(inoperative) (operative)

slow Ca2+channel in myocardial cells

Ion channels can be regulated by G-proteins !
(the muscarinic acetylcholine receptor)
release of Ach by nerve fibers 

binding of Ach to G-protein-linked

receptor in heart muscle cells 

G-protein activated, GDP-GTP

exchange, dissociation 

binding of -complex
to K+ channel 

opening of channel, outward flow of

K+  hyperpolarization  slowing of
heart beat 

hydrolysis of GTP, - reassociation,

inactive G-protein, channel closing
MCB. Ch13 p566
IV. Physiological functions of ion
channels
 REVISION
Major physiological functions of ion channels

Charge transfer determination of the membrane potential

resting potential
RT pK [K]i  p Na [Na]i  pCl [Cl] o
Em   ln
Fz pK [K] o  p Na [Na] o  pCl [Cl]i

extracell.

cytosolic
K+ Na+ Cl−
 REVISION
Major physiological functions of ion channels

Charge transfer determination of the membrane potential

+50
resting potential

Membrane potential (mV)

rising phase:
depolarization
action potential and 0
declining phase:
synaptic potential repolarization

threshold sub- hiperpolarizing after

threshold potential
−70
stimuli

conductivity [mmho/(cm2)]

Na+ permeability

K+ permeability

time (ms)
 REVISION
Major physiological functions of ion channels

Charge transfer determination of the membrane potential

RT pK [K]i  p Na [Na]i  pCl [Cl] o
resting potential Em   ln
Fz pK [K] o  p Na [Na] o  pCl [Cl]i
action potential and
extracell.
synaptic potential
cytosolic
K+ Na+ Cl−

Ion transferchanging cytosolic ion concentrations

A. neural
rise of [Ca2+]i 1
action potential

2
3 5
4

Muscle cell

MBC. Ch11 p687 Sarcoplasmic reticulum

 Major physiological functions of ion channels !

Charge transfer, electric signal (action pot. and synaptic pot.):

Voltage-gated K+ and Na+ channel
and ligand-gated nAChR

Ion transfer, channels influencing cytosolic Ca2+

concentration
Ca2+ channels

plasma membrane intracellular membrane

Voltage-gated: CaV1.2
-RyR
[Ca2+]=2 mM -IP3R
Ligand-gated: P2X [Ca2+]=~ mM MBC. Ch11 p688
[Ca2+]=100 nM MBC. Ch15 p912
Events during synaptic transmission-chemical synapse
!

action potential in presynaptic cell  depolarization  opening of voltage-gated

Ca2+ channels  exocytosis of ACh containing vesicles  diffusion of the
neurotransmitter (ACh) across the synaptic cleft  binding of ACh to receptors
 opening of cation channel  depolarization of postsynaptic cell  action
potential
MBC. Ch11 p687 MCB. Ch7.8 p290
 The nicotinic acetylcholine receptor: a key player of synaptic
transmission in the neuro-muscular junction (NJM)
!
Selects based on size and charge (ligand-gated
cation channel, mildly selective, but mostly
allows Na+ influx, so depolarizes)

Binding of two acetylcholine molecules opens

the channel to allow the passage of cations.
filter
Agonists: ACh and nicotine
(bind to and activate the channel)
Antagonist: curare (Indian arrow poison)
(binds to, but does not activate the channel,
competes with ACh, breathing muscles unable
to contract)

Channelopathy:

MBC. Ch11 p684 myasthenia gravis (skeletal muscle weakness)

MCB. Ch7.8 p292 info autoantibodies against nAChR (loss of nAChR)
MBC. Ch11 p682
Synapses can be excitatory or inhibitory !
Extracellular Extracellular
space space
Glutamate GABAA
receptor receptor
Intracellular Intracellular
space space
_
Na+ Cl

-55 mV AP threshold -55 mV AP threshold

-65 mV -65 mV
-70 mV
Sub-threshold
Excitatory Postsynaptic Inhibitory Postsynaptic Potential
Potential (EPSP) (IPSP)
higher chance of action potential lower chance of action potential
Neurotransmitter receptors, that are ligand-gated ion channels
Functional type ligand Ion selectivity
Excitatory receptors Acetylcholine (nicotinic) Na+ / K+
glutamate (NMDA-type) Na+ / K+ and Ca2+
glutamate (non-NMDA type) Na+ / K+
serotonin Na+ / K+
Inhibitory receptors GABA Cl-
glycine Cl-
 Ionotropic versus metabotropic receptors !
Example 1:
nAChR vs. mAChR
Same signal molecule,
different receptor
different mechanism
different effect

Ionotropic receptor: Metabotropic receptor:

ligand gated ion channel – the signal
molecule binds to the ion channel itself
Fast action – short effect
Example: nicotinic Ach receptor
Cation channel, depolarization
Not an ion channel itself - the signal
molecule binds to the receptor and the
signal is transmitted via a pathway
(often through a G-protein) to an ion
channel
Slow action – longer lasting effect
Example: muscarinic Ach receptor
mAChR--»G-protein--»βγ--»
K+ channel opening--»hyperpolarization
Ionotropic (iGluR) vs. metabotropic (mGluR) glutamate info
receptors -- same signaling molecule -- different effects

iGluR mGluR

dark
iGluR

Extracellular
space

Intracellular
space
Na+
light
Extracellular
space

Intracellular Na+/Ca2+
space
!
 Electric versus chemical synapses
!
Electric synapse Chemical synapse

direct flow of ions via gap junctions transmission via neurotransmitters

advantage: advantage:
nearly instantaneous (cardiac muscle) -signal amplification ( 1 motor neuron
 contraction of multiple muscle
cells)

- computation (Neurons form a huge

network of many connections,
receive signals at multiple excitatory
and inhibitory synapses. They
combine, interpret and record these
signals, which forms the basis for
acting, thinking, perceiving and
remembering.)
V. Ion Channels and Disease
The shape of action potentials is determined by REVISION
ionic currents

Delayed opening of K+ channels in

shaker mutant Drosophila motor
neurons

Ionic currents contributing

to the shape of cardiac
action potentials
Long QT syndrome
1. QT interval > 450 ms
!
2. torsades de pointes
(rapid contractions, reduced
ventricular refilling, lower cardiac
output, less blood flow to the brain)
3. fibrillation
……sudden death

3
 !
Increase of sustained INa or decrease in IK prolongs action
potential duration and QT interval (long QT syndrome)

INa
+40 mV IKs
IKr
INa

-80 mV
200 ms
R
P

Q S T Q T
 Mutation in the KATP channel results in neonatal diabetes !
Low glucose concentration High plasma glucose Mutation in KATP channel

No insulin
Insulin No insulin
secretion Glucose Glucose
release secretion

Metabolism Metabolism
Metabolism

ATP ATP
ATP
MgADP
MgADP MgADP
Ca2+

-70 mV

Hiperpolarized Hiperpolarized
K+ membrane Depolarization Ca2+ K+ membrane
Open KATP Closed Ca2+ Closed KATP Open Ca2+ KATP channels Closed Ca2+
channels channels channel channels cannot be blocked channels
by ATP, therefore
they are open

Fasting: low metabolism, Food intake: increased metabolism Food intake: increased metabolism
KATP channels are open KATP channels are closed KATP channels are open
minimal insulin secretion increased insulin secretion Minimal insulin secretion

Neonatal diabetes
 Channelopathies
Autoimmune diseases attacking ion channels – Myasthenia gravis (AchR),
Lambert-Eaton syndrome (skeletal muscle Ca2+ channel)

Myotonia (extreme muscle tension) – skeletal muscle Na+ channels do not

fully inactivate, which results in enhanced muscle excitability

Long QT syndrome – cardiac Na+ and K+ channels

Malignant hyperthermia – anesthesia  rise in [Ca2+]i in skeletal muscle 

strong contraction  potentially fatal rise in body temperature

Cystic fibrosis – mutation in a Cl- channel  reduced epithelial Cl-

conductance  mucous accumulation in the lungs  chronic infection 
death

Startle disease (hyperekplexia) – mutation of the glycine receptor in the

CNS; muscle spasm in response to an unexpected stimulus, facial grimacing,
clenching of fists, jerking of limbs, the whole body becomes rigid
VI. Ion Channels as Targets for Drugs
 Ion channels as targets for drugs I.
!

Local anesthetics (e.g. lidocaine) block Na+ channels:

• local injection in high concentration blocks impulse
conduction in nerves
• systematic application in low concentration – antiarrhythmic
action

Ca2+ channel blockers (verapamil, diltiazem, nifedipine):

• antiarrhythmic action (verapamil, diltiazem)
• vasodilator action (hypertension)

!
 Ion channels as targets for drugs II.
!

Sedatives, antianxiety agents:

• These stimulate the inhibitory synapses of the CNS; GABA
receptors open more frequently (benzodiazepines, Valium,
Librium) or stay open for longer durations (barbiturates) –
hyperpolarization of neurons, depression of CNS, sedating
effect

!
 info
Ion channels as targets for drugs III.

Lymphocyte K+ channel blockers as potential immunosuppressants:

(work at the electrophysiology lab of the Department)

• certain T lymphocytes (effector memory cells) that are

involved in the pathogenesis of autoimmune diseases (e. g.
multiple sclerosis, type I diabetes) express very high numbers
of a K+ selective ion channel. If these channels are blocked, the
activity and proliferation of these autoreactive T cells are
suppressed and the symptoms of the disease are relieved.