KV1.3 | Shaker Related Potassium Channel Member 3

Family:
Potassium channels

Subgroups:
Shaker (KV1.1–KV1.8), Shab (KV2.1-KV2.2), Shaw (KV3.1–KV3.4), Shal (KV4.1–KV4.3), KQT like (KV7.1–KV7.5), Eag related (KV10.1-KV10.2), Erg related (KV11.1–KV11.3), Elk related (KV12.1)

Topology:
Contains six transmembrane domains (S1–S6), four single subunits form a pore, homotetramers and heterotetramers are possible.

KV1.3 Background Information

KV1.3 belongs to the delayed rectifier class, members of which allow nerve cells to efficiently repolarize following an action potential. KV1.3 is expressed in T and B lymphocytes and plays an essential role in T cell proliferation and activation. Blockade of KV1.3 channels in effector-memory T cells suppresses calcium signaling, cytokine production (interferon-gamma, interleukin 2) and cell proliferation. KV1.3 is an important target for drug development for autoimmune diseases as Multiple Sclerosis, type-1 diabetes mellitus and rheumatoid arthritis.

Gene:
KCNA3

Human Protein:
UniProt P22001

Tissue:
brain, lung, osteoclasts, T-lymphocytes, B-lymphocytes

Function/ Application:
T-lymphocyte activation, apoptosis, proliferation

Pathology:
Immune response, multiple sclerosis, rheumatoid arthritis, diabetes mellitus, asthma, cancer

Interaction:
KVβ2, β1 Integrin, SAP97, ZIP

Modulator:
PAP-1, Margatoxin, Noxiustoxin, Charybdotoxin

Assays:
Patch Clamp: whole cell, room temperature, physiological temperature, State- and use-dependence / site of action

Recommended Reviews:
Gutman et al. (2005) International Union of Pharmacology. LIII. Nomenclature and molecular relationships of voltage-gated potassium channels. Pharmacol Rev 57(4):473-508

Data and Applications

KV1.3 - Colour Coding Displays Real Gigaohm Seals

0Kv13icon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Evotec AG, Hamburg, Germany.

Shown is a screenshot made during a recording of KV1.3 currents stably expressed in CHO cells. On the top left the raw currents from each individual cell are displayed. The background color coding marks the cells with seals above 500 MΩ in blue and above 1 GΩ in green.

KV1.3 - Continuous Internal Perfusion

p24 1 internalPerf

icon pap   Port-a-Patch data and applications:

KV1.3 currents (blue), endogenously expressed in Jurkat cells, were rapidly blocked by internal perfusion of Cs+ (light blue), and fully recovered after washout with K+ (grey). Internal solution replacement was repeated 19 times and the recording was stable for over 35 minutes, as shown in the lower graph.

KV1.3 - Internal Application of TEA and Quinidine

icon pap   Port-a-Patch data and applications:p25 1 Kv1,3

KV1.3 current was blocked by the internal application of increasing concentrations of quinidine (left) or TEA+ (right). For quinidine the IC50 was determined as 15 μM (literature value external application 14 μM) and for TEA+ the IC50 was determined as 0.9 ± 0.3 mM (n = 3) (literature value internal application 0.6 mM).

KV1.3 - Internal Perfusion

p41 1 IntPerficon sp96   SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:

The SyncroPatch 96 supports internal perfusion allowing internal administration of compounds, second messengers and metabolites. Here, KV1.3 currents, endogenously expressed in Jurkat cells, were blocked by the internal administration of Cs+ followed by washout with Cs+-free internal solution.

KV1.3 - Internal Solution Exchange during recording

p35 4 ExchInternSolicon pl   Patchliner data and applications:

A unique feature of the Patchliner is its ability to exchange the internal solution during the recording. The figure shows recordings of KV1.3 from two Jurkat cells (simultaneoulsy recorded) in the presence of control internal solution, after the exchange of the internal solution with a Cs+ solution, and subsequent washout (left to right).

KV1.3 - Perforated Patch

application hkv13 1application hkv13 2

icon pap   Port-a-Patch data and applications:

Perforated and conventional whole cell configuration derived hKV1.3-currents. Voltage-dependence was shifted significantly to more negative potentials in the whole cell configuration compared to the perforated whole cell configuration. Whole cell currents after a conventional membrane breakthrough (right) and perforated whole cell currents (left).

KV1.3 - Pharmacology with High Success Rate

Kv13 384 raw onl norm Quini 2icon sp96   SyncroPatch 384PE data and applications:
Cells were kindly provided by Evotec.

Shown are screenshots of a pharmacology experiment performed with the SyncroPatch 384PE. Recordings from 384 KV1.3 expressing CHO cells were performed simultaneously. Original current traces and the peak current over time are displayed. Data are analysed with DataControl384 full analysis tool. With just a few mouse-clicks normalized concentration response curves can be generated. Here, normalized response and the IC50 of Quinidine is shown. Darkening shades of blue indicate increasing compound concentration.

KV1.3 - Reproducible Compound Application

icon pl   Patchliner data and applications:p36 1 reproducRec

Application of 5 μM quinidine leads to about 50 % block of the KV1.3 currents (blue). After washout, the current is fully recovered (grey). The lower graph shows corresponding Imax (+40 mV) in the absence and presence of 5 μM quinidine, for two different cells with eight consecutive application and washout steps. The recording lasted over 40 minutes!

KV1.3 - Voltage Induced Membrane Movements Measured with Atomic Force Microscopy

icon pap   Port-a-Patch data and applications:Porti AFM JurkatWebData are taken from Pamir E., et al., Ultramicroscopy, 2008, 108, 552-557.

Voltage induced membrane movements of a Jurkat cell. (a) Deflection signal of the cantilever resting on the cell membrane at an indenting force of 1.0 nN. (b) Corresponding whole cell current: The characteristic response of the voltage gated potassium channel KV1.3 in Jurkat cells is observed. (c) Below: Applied pulse protocol.

Application Notes

Publications

2007 - Automated ion channel screening: patch clamping made easy

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Expert Opinion Therapeutic Targets (2007)

2008 - Planar patch-clamp force microscopy on living cells

icon pap  Port-a-Patch publication in Ultramicroscopy (2008)

2008 - Synthesis and biological evaluation of chalcones as inhibitors of the voltage-gated potassium channel Kv1.3

icon pap  Port-a-Patch publication in Bioorganic & Medicinal Chemistry Letters (2008)

2009 - Port-a-Patch and Patchliner: High fidelity electrophysiology for secondary screening and safety pharmacology

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Combinatorial Chemistry & High Throughput Screening (2009)

2016 - Human T cells in silico: Modelling their electrophysiological behaviour in health and disease

icon pl  Patchliner publication in Journal of Theoretical Biology (2016)

2017 - High-throughput electrophysiological assays for voltage gated ion channels using SyncroPatch 768PE

icon sp96  SyncroPatch 768PE publication in PLoS One (2017)

2017 - Potassium channels Kv1.3 and KCa3.1 cooperatively and compensatorily regulate antigen-specific memory T cell functions

icon sp96  SyncroPatch 768PE publication in Nature Communications (2017)

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