Structural and Functional Diversity of Several Ion Channels in Cardiac Tissue

Introduction
Ion channels are at the core of every physiological function. They allow cells to maintain their integrity
through mechanisms such as volume regulation. They can also create electrical signals which enable
all of our motor and sensory functions. These functions allow humans to perceive and operate in the
world around them.

Experimental techniques such as electrophysiological recordings have enabled us to have a better

insight into the properties of these membrane channels. Despite being similar in structure, ion
channels can have variable and complex properties that result in functional diversity.

The heart pumps blood around the body by constantly beating rhythmically throughout a person’s
lifespan. This rhythm is controlled autonomously via a series of electrical signals regulated by
pacemaker cells. These pacemaker cells are located at the sinoatrial node, atrioventricular node and
along the septum as the bundle of his and Purkinje fibres (Unudurthi, Wolf and Hund, 2014). The heart
propagates those regulated electrical signals using electrical synapses between cardiomyocytes
(https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3210376/).

The present study will discuss the structural and functional diversity of the calcium-activated
potassium (KCa) channels, Two P domain potassium (K2P) channels, Voltage-gated potassium (Kv)
channels and Inwardly rectifying (Kir) potassium channels. The functional diversity will be discussed in
relevance to ventricular cardiomyocytes and sometimes the cardiovascular system.

Structural comparison of different ion channel protein classes

Nav and Cav both contain a single α subunit containing four separate transmembrane regions all of
which are part of the same polypeptide. Each transmembrane region is identical to the others and
spans the membrane six times and contains a single pore-forming region between the 5th and 6th
transmembrane domain. The transmembrane domains are made up of β pleated sheets while the rest
of the protein consists mainly of α helices. The difference between Nav or Cav and potassium channels
is that instead of having a single subunit with four domains, the Kv channels have four separate α
subunits each containing a single transmembrane domain. Similarly, each subunit of the Kv channels
contains six transmembrane regions with a pore-forming region between the 5th and 6th
transmembrane region. Any single polypeptide subunit such as the single α unit in the Nav or Cav or
any of the four separate α subunits in the Kv channels has an intracellular N-terminal and C-terminal.
Nav, Cav or Kv channels can all have optional β auxiliary subunit for a variety of different functions
such as “ball and chain” inactivation where the cytoplasmic n-terminal β subunit covers the pore of the
channel preventing flux. Additionally, Cav can also have α2δ and γ subunits to enhance surface
expression or for other functions (Marban, Yamagishi and Tomaselli, 1998) (Catterall, 2011).

The KCa, Kir or K2P channels can differ more in structure. There are three different types of KCa
channels in humans characterised by different channel conductances. Those are the big conductance
Kca (BK) channels, intermediate conductance Kca (IK) channels and the small conductance Kca (SK)
channels. The IK and SK are sometimes both considered SK channels due to how closely related they
are.
SK and IK channels share the same α subunit structure as the Kv channel, also containing a voltage
sensor on the 4th domain and a pore-forming region. In contrast, BK channels significantly differ in
structure. The N-terminus of the α subunit is present in the extracellular matrix, and the polypeptide
spans the membrane seven times before extending into the cytoplasmic space to form four more
domains for a total of eleven domains ending at the C-terminal inside the cytoplasmic space. The
pore-forming region is present between the 6th and 7th transmembrane regions. Unlike BK channels,
SK and IK channels sense Ca2+ using a c-terminal intracellular messenger protein known as
calmodulin.

Lastly, Kir and K2P share a similar structure. For the Kir channel, the α subunit contains only two
domains and a single pore-forming region starting at the cytoplasmic N-terminal and ending at the
cytoplasmic C-terminal. For the K2P, each α subunit contains four transmembrane domains and two
pore-forming regions. The structure is equivalent to two Kir channels attacked intracellularly with one
another between the second transmembrane domain of one channel and the first transmembrane
domain of another.

All the channels described can have auxiliary subunits that supplement their properties and functions.
(Kuang, Purhonen and Hebert, 2015).

Comparison of different ion channels function in the cardiovascular system

Sodium channels

Nav1.5 channels are the most expressed channel in cardiomyocytes. Following pacemaker activity,
cardiomyocytes receive an electric current that depolarizes the cell past the threshold voltage for Nav
channel activation. This causes Nav channels to open allowing a strong inward current of cations that
rapidly depolarizes the cell (DeMarco and Clancy, 2016). Nav1.5 channels close after brief activation
through a rapid C-terminus inactivation mechanism known as the “ball and chain” (Savio-Galimberti,
Gollob and Darbar, 2012).

Calcium channels

In comparison to the Nav channel, the Cav channels play a much more complex role in excitable cells.
Cells normally have a minute intracellular calcium concentration of about 8x10-5 mM to prevent a
reaction with phosphate that forms harmful insoluble aggregates (Niederer, 2013).

There are four main types of Cav channels characterised by different voltage, 1,4-dihydropyridine, ω-
agatoxin and ω-conotoxin sensitivities. Those are known as the L, N, P/Q or R type Cav channels.

The L-type Cav 1.2 channels are responsible for the upstroke depolarization potential of sinoatrial
node pacemaker cells. Following late phase diastolic depolarization (pacemaker phase), the voltage
threshold for channel activation is reached causing channel activation and cation influx into the cell
initiating the next action potential. The Ca2+ influx then activates ryanodine receptors 2 which are
responsible for store-operated calcium release from the sarcoplasmic reticulum (Yamakage and
Namiki, 2002) (Mangoni et al., 2006).

Potassium channels

Channels for potassium ions have a lot more variation in their function depending on the potassium
channel type and the excitable cell it is present within. These often play a role in establishing a resting
membrane potential through different mechanisms that in coordination with other channels give rise to
the characteristic cardiac action potential. However, their functions aren’t limited to just establishing a
resting membrane potential.

Kv channels activate in response to voltage changes and are involved at many stages the cardiac
action potential. These are functionally categorized into six categories: Delayed rectifying, outward
rectifying, inward rectifying, slowly activating, A-type and modifiers. Following the depolarization of
cardiomyocytes, The Shaker and Shal related A-type Kv channels allow a transient outward K+ current.
This causes a slight decrease in membrane potential before rapid N-terminus or else inactivation
(Pongs, 1993) (Wollberg and Bähring, 2016). During the stationary phase, delayed rectifying Kv
channels activate and allow an outward K+ ion flow that opposes the Ca2+ influx resulting in a
somewhat neutral net charge exchange across the membrane. When the Cav channels close, delayed
rectifying Kv channels remain open causing the initial net positive current that initiates
hyperpolarization (Jeevaratnam et al., 2017). The reduction in membrane potential causes the
activation of inwardly rectifying “ether-à-go-go" Kv11.1 channels and Kir channels. These speed up
repolarization to achieve a resting membrane potential at which delayed rectifying channels deactivate
(Sanguinetti and Tristani-Firouzi, 2006).

The Kir channels are comprised of the classical Kir, g protein-gated Kir and ATP sensitive Kir channels.
In cardiomyocytes, classical Kir channels are responsible for establishing a resting membrane potential
by allowing K+ efflux at rest. However, upon depolarization, this outward current gradually decreases
until the channel becomes virtually closed. This prevents cardiomyocytes from directly initiating the
repolarization phase and allowing the stationary phase of the cardiac action potential to occur. When
repolarization initiates by Kv channels the voltage decreases allowing classical Kir channels to open
and aid in accelerating repolarization. (Hibino et al., 2010). G-protein-gated Kir channels are coupled to
acetylcholine sensitive GPCRs which enable parasympathetic modulation. Acetylcholine causes
channel activation that leads to hyperpolarization of the cardiomyocyte. (Krapivinsky et al., 1995). ATP
sensitive Kir channels are made up of four ATP sensing Kir6.1 α-subunits and four ADP sensing
sulfonylurea subunits. Their function is to respond to a reduction in the ATP:ADP ratio during
ischaemia by opening. This characteristic hyperpolarization enables a mechanism known as
ischaemic preconditioning which plays an important role in preventing severe ischaemic damage
during a myocardial infarction (Hibino et al., 2010).

Kca channels detect intracellular voltage or calcium ions to activate (Wang et al., 1996). These
channels present the least relevance in cardiac function. However, they do have novel functions in
cardiac tissue and cardiovascular, endothelial tissue. BK channels have a conductance in the
approximate range of 100-300 pS (Lee and Cui, 2010). BK channels are distinguished from other Kca
channels in their ability to activate at physiologically irrelevant voltages as well as Ca2+ or Mg2+. BK
channels on the mitochondrial membrane of cardiomyocytes have been found to play a protective role
in reducing ischaemic damage during a myocardial infarction (Hermann, Sitdikova and Weiger, 2015).
The IK channels have a conductance in the approximate range of 20-85pS (Ishii et al., 1997). IK
channels are found on cardiac fibroblasts where they play a role in proliferation. They also play a role
in regulating pacemaker activity (Dong, Bai and Cai, 2016). Similarly, IK channels also show
expression on endothelial cells where they cause hyperpolarization that propagates to the vascular
smooth myocytes leading to vasodilation and a reduction in blood pressure. The SK channels have a
conductance in the approximate range of 2–20pS (Ishii et al., 1997). They are expressed in
cardiomyocytes and activated through a ryanodine receptor-mediated Ca2+ influx which occurs during
depolarization events such as ventricular fibrillation. SK channels function to hyperpolarize and
reverse such events to regulate the action potentials and ultimately the contraction of the heart
(Diness et al., 2015).

Unlike other channels, K2P channels are insensitive to voltage change and many channel blockers.
This allows them to influence the resting membrane potential time independently and without
responding to changes in ion concentrations during polarisation events. Different K2P types can be
sensitive to pH, lipids or mechanical stretch depending on their function (Tamargo, 2004).

Discussion

The ion channels reviewed in this paper share a recurring structural theme that enables them to
respond to signals and conduct ions through the plasma membrane. This involves single or multiple
polypeptide chains that span the membrane. Voltage sensing ion channels share a similar
transmembrane domain in their structure. Similarly to voltage sensing channels, ion sensitivity is
enabled through ion sensing structures which are part of the channel protein. The structural variations
amongst these channels allow them to be responsive or unresponsive to certain events. The overall
harmony gives rise to a characteristic electrical profile that is identical throughout electrically coupled
cardiomyocytes and regulated by pacemaker cells that have a different ion channel composition.
Overall, these properties allow the maintenance of lifelong synchronised contractions and relaxations
of cardiomyocytes.

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