I.

Introduction

Voltage gated potassium channels are one member of the family of ion channel proteins. These

proteins consist of pores in the cell wall gated by an action potential. Once the action potential is

triggered, the pore opens and ions are allowed to diffuse through the membrane. Voltage gated

potassium channels are extremely rapid in diffusion while also being extremely selective. This,

combined with the fact that the structure and mechanism of this protein is conserved even between

bacteria and humans, indicates that there is practically no room for improvement. The mechanism is

ideal for its purpose.1

The process of voltage dependent ion diffusion was first studied in 1954 by Hodgkin and Huxley

who suggested that the mechanism involved a single-file stream of ions through the membrane. Once

this process was linked to voltage gated potassium channels, they have been most extensively studied

in Drosophila melanogaster. The Shaker potassium channel was sequenced in 1987, beginning a more

complete of investigation into the protein family. Roderick MacKinnon won the Chemistry Nobel Prize

in 2003 for his work with voltage gated potassium channels determining the atomic basis of selective

ion conduction.1,2,3

This protein family shows great promise in the development of pharmaceuticals. Ionic

concentration and flow is involved in a huge number of cell functions in humans. Voltage gated

potassium channels play a role in regulating muscle function, cell proliferation, neural signaling and

apoptosis. However, this diversity of function in the body creates a new problem. There is a high

potential for side effects due to the number of very similar proteins. As will be discussed later, voltage

gated potassium channels are heterotetramers made up of subunits that vary significantly in the non-

essential sequences. Drugs can take advantage of this by targeting only the specific subunits, or
combinations of subunits, that are present in the target cells without interfering with other, similar,

channels.4,5,6

II. Biological Function

Voltage gated potassium channels serve to regulate membrane potential and ionic concentration.

Opening the pore results in the free movement of positively charged potassium ions across the

membrane. This allows electrical energy to flow and ionic concentration to be regulated. This drives

the mechanism behind action potentials as well as contributing to apoptosis and cell proliferation.5

Action potentials are what allows neurons to fire and muscle cells to contract. At rest, a strong

ionic gradient is maintained at the cell membrane; the inside is negatively charged while the outside is

positively charged. This changes as the muscle contracts or neuron fires. Sodium ion channels are

opened first and sodium ions to pour into the cell due to osmotic pressure. This leads to a change in

the voltage of the cell membrane, causing voltage gated potassium channels to open. This accelerates

the change in the charge to its maximum speed, before it reaches the tipping point. A balance is

reached and any excess ions start to flow in reverse. Afterwards ionic pumps are used to remove

enough of the remaining ions to reset the action potential.5,7

Variations on voltage gated potassium channels can contribute to modulating the intensity,

duration and frequency of cell actions. This is the sometimes the source of the disease arrhythmia. In

arrhythmia, the muscles in the heart have flawed electrical signaling, which causes abnormal rates of

contraction. Research has been done into anti-arrhythmic drugs targeting the cardiac voltage gated

potassium channels. Both blockers and openers can serve to help regulate cardiac function and return

it to healthy levels depending on the case in question.5,4

Similar principles apply to neural signaling. Conditions such as epilepsy and Alzheimers involve

levels of neural excitation that are too high, too low or even non-existent in certain areas of the brain.
Regulating the voltage gated potassium channels can help return neural excitation to healthy levels. In

this area, the primary challenge in designing drugs is specificity; the drugs must be able to interact with

very specific varieties of the ion channels to prevent side effects.5 4

The role of voltage gated potassium channels in cell proliferation is very similar to its role in action

potentials, though reversed. T-Lymphocytes and prostate cancer cells, among others, rely on an influx

of calcium ions to trigger cell proliferation and, just as in action potentials, this process results in a

positive charge within the cell that polarizes the membrane. The charge triggers the voltage gated

potassium channels which, rather than allowing potassium into the cell, release it from the cell to

balance the charge and encourage the continuing necessary flow of calcium ions.5,4

The application of this information to drug development is quite straightforward. Blocking or

otherwise inhibiting the channels on T-lymphocytes can serve as an immunosuppressant, while the

value of suppressing cancer cell proliferation is self-evident.

Apoptosis, or programmed cell death, is not as well understood as the other processes discussed

here and consequently the role of voltage gated potassium channels in it is also not as well

understood. Regardless, there are several steps of the process that can be linked to voltage gated

potassium channels. A common characteristic of apoptosis is a simultaneous decrease in cell volume

and efflux of potassium ions into the cell. The cause of the volume decrease is the release of water

and other cations from the cell, which occurs at roughly the same time as nuclear breakdown and DNA

fragmentation. Consequently, the high potassium concentration results in a hyperpolarization

between the cell and the mitochondria more than twice as powerful as the polarization in action

potentials. This hyperpolarization has been linked to the release of cytochrome c and the death of the

mitochondria prior to the overall death of the cell, but the exact cause is yet to be determined.8 6
 It is of some interest that the effect of drugs on cell apoptosis is very inconsistent. Drugs targeted

at inhibiting the subunits of the channels involved will encourage apoptosis in some cells while

inhibiting it in other cells. This has been proposed as a method for combatting cancer cells but further

research is required to understand the process.6

III. Structure, Mechanism and Chemistry of Function

There are two main components to a voltage gated potassium channel: the voltage sensor and the

ion conduction pore. The voltage sensor must be able to detect a change in charge on one side of the

membrane and then respond by causing a conformational change in the protein that will open the

pore. The ion pore must be both highly selective and extremely fast. Once opened, it must be able to

allow only potassium ions through while excluding similar molecules like sodium ions. It must be able

to allow these molecules through with a very low energy cost so they can flow with a rate high enough

to trigger action potentials. These two components are what allow voltage gated potassium channels

to effectively serve their role in cells.

Voltage gated potassium channels are made up of complexes of four subunits. Some channels are

homotetramers while others are heterotetramers. This paper will focus on the KvAP protein found in

Aeropyrum pernix, a member of the phylum archaea, which is made up of homotetramers.9 Nobel Prize

winner Roderick MacKinnon used KvAP in his research due to its high degree of similarity to Shaker:

the most documented family of voltage gated potassium channels.1 The Shaker family is present in

insects, mammals and plants, while its cousins, including KvAP, are present in bacteria.10,11

Each tetramer of KvAP is made up of a series of 6 subunits, S1-S6, each of which contains one alpha

helix, save S3 which contains a and b helixes. The voltage sensing region is located on the paddle

made up of subunits S1-S4. There are four arginine residues in the S3-S4 area which are responsible

for triggering the movement of the paddle (See Figure 1).12 Conventional models indicated that the
movement of the charges was entirely within the greater structure of the protein, but the first X-ray

crystallography of KvAP by Jiang et at in 2003 suggested a more direct method.13,9 According to his

model, the paddle moves through the entire membrane, hinged by glycine residues at the S4-S5

linkage, to bring the gating charges to rest in the external solution (See Figure 2).14 Each subunit of the

tetramer moves in tandem to cause the large scale conformational change which opens the pore. This

is the physical mechanism behind the response of KvAP to voltage.

The function of the second component of the voltage gated potassium channel, the ion conduction

pore, is far less kinetic and more chemical. The pore must allow only potassium ions through while

excluding similar molecules like sodium. Experimentally, it has been determined that potassium

channels select for potassium over sodium by a factor of 1000, while operating at an extraordinarily

high rate of 108-109.15 The rate at which potassium channels operate is close to diffusion, while also

being highly selective.

The selectivity of potassium channels is dependent upon one extraordinarily conserved sequence:

TVGYGD. This sequence is found in voltage gated potassium channels in mammals and bacteria, as

well as being found in other channels like calcium gated potassium channels (See Figure 3). We can

deduce that this function must be extremely important to the conduction of potassium. This sequence

is located on a loop connected to the S6 region (See Figure 1).9 The four subunits fit together so that

four parallel copies of this sequence line the pore. The mechanism by which this sequence

discriminates was discovered by Roderick Mackinnon. The dihedral angle of the two glycine residues,

the coordination from the partial negative of threonine and the structural support of tryptophan and

valine in in the hydrophobic core allow the backbone of the peptide chain to form coordinating bonds

ideal for potassium.1 The end result is four evenly spaced binding sites for potassium within the pore.

Each binding site is quite similar to how a potassium ion would be kept in solution: a series of eight
oxygen-potassium bonds forming a twisted cube around the atom. The oxygens are all held in place by

the geometry of the conserved sequence so, unlike water, it cannot adjust the length of the bonds to

accommodate a similar atom like sodium.1 Up to two potassium ions are found in the pore at any one

time, spaced out by water molecules.16 The importance of these water molecules is based in the other

important feature of the pore: its energetic favorability.

Voltage gated potassium channels operate at approximately the rate of diffusion.15 Ions are

passing through the channel very nearly as quickly as if there were no pore at all and it was just an

aqueous channel through hole in the membrane. This is as fast as it is physically possible for this

process to occur.

There are two potential energetic costs to the conduction of potassium: pulling potassium ions out

of aqueous solution and breaking bonds with the pore to release the ion once it passes through the

pore. Each of these potential costs are minimized by individual mechanisms.1 Firstly, since

dehydrating potassium ions to bring them into the pore would cost enough energy that it would

significantly slow the flow of ions, to maintain the flow,the channel simply doesnt dehydrate them.

While there are four binding sites in the channel, only two are occupied by potassium at any given

time.17 The flow rate through the pore is consistent with an average occupancy of 0.5 per binding

site.18 The other two binding sites are occupied by water. Water and potassium alternate to maintain

hydration within the pore and complete the ideal molecular geometry for potassium selectivity. 1 The

bond breaking challenge is overcome through a similarly elegant mechanism. Since energy is released

as the bonds form, it must be added to break the bonds on the other end of the pore; the channel

effectively recycles this energy to overcome the energy cost of release. If only one potassium ion is

within the channel instead of two, then the end of the channel collapses and the flow will not continue
until a second potassium ion starts to bind and pushes the first ion through and out of the pore. 16

These two combine to allow the free and easy flow of potassium through the pore.

The mechanism of the voltage gated potassium channel is straightforward, but it is so well adapted

to its role that it functions very nearly as well is physically possible, with nearly ideal selectivity and

speed.

IV. Conclusion

In terms of function and structure, voltage gated potassium channels are exceptionally well

understood. The mechanisms of voltage sensing, pore opening and selectivity have all been studied

extensively and are well documented. However, the part it plays in the overall function of various cells

remains an important area for research. Targeting specific potassium channels with pharamceuticals

could allow the regulation of a wide variety of cell functions, such as muscle contraction, brain

function, cell proliferation and apoptosis; provided the role of voltage gated potassium channels in

these processes can be more precisely determined.

Figure 1:
a. b.

Crystal structure of KvAP: a. One subunit of the KvAP channel. In red: arginine residues 117, 120, 123, 126
responsible for voltage sensing. In green: Highly conserved potassium pore selectivity filter sequence: TVGYGD. b.
1ORQ full tetramer. In green: Antibodies used in the crystallization process, not part of the protein. Image
adapted from paper by Jiang Et al.
Figure 2:

K+ K+

+ +

+ +

Figure 2: Mechanism of Ion pore opening

The paddles swing through the membrane to move the signaling charges to the outside of the
cell. In the process a conformational change opens the ion pore.

Figure 3:
Mammalian Shaker 361 VYFAEADERD-SQFPSIPDAFWWAVVSMTTVGYGDMVPTTIGGKIVGSLCAIAGVLTIAL
Streptomyces Kcsa 24 LAVLAERGAPGAQLITYPRALWWSVETATTVGYGDLYPVTLWGRCVAVVVMVAGITSFGL
Calcium gated MthK 27 GFHFIEGE-------SWTVSLYWTFVTIATVGYGDYSPHTPLGMYFTCTLIVLGIGTFAV

Potassium Selectivity Sequence

The potassium selectivity sequence TVGYGD is conserved in both a mammalian Shaker
family voltage gated channel, the bacterial Kcsa voltage gated channel as well as the
calcium gated potassium channel MthK.
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