Kubota T , Correa AM , Bezanilla F .

R01 GM030376 NIGMS NIH HHS , U54 GM087519 NIGMS NIH HHS

	Figure 1. N-terminus of Kv1.3 prevents the expression on Xenopus oocytes membrane.(A) Amino acid sequence alignments between human Kv1.3 (Kv1.3), human Kv1.2 (Kv1.2) and Shaker K channel (Shak). Two potential start codons are highlighted in red circles in Kv1.3. The N-terminal sequences in Shaker K channel responsible for N-type inactivation is highlighted in red in Shak. The T1 domain is highlighted in light yellow. (B,C) Representative gating currents from hKv1.3-W436F (B) and from hKv1.3-del-W436F (C). (D) Normalized charge as a function of voltage (Q-V) for hKv1.3-W436F (open circles, n = 4) and for hKv1.3-del-W436F (filled circles, n = 8). Error bars indicate standard error of mean (SEM). (E) The total charge as a function of fluorescent intensity for hKv1.3-W436F-mCherry (open circles) and for hKv1.3-del-W436F-mCherry (filled circles). These data were obtained from the same batch of Xenopus oocytes. Each data set was fitted to a linear function (dotted lines). The R2 values were 0.98 for hKv1.3-del-W436F-mCherry and 0.91 for hKv1.3-W436F-mCherry. The linear slopes were 3.4 ± 0.2 for hKv1.3-del-W436F-mCherry and 1.1 ± 0.2 for hKv1.3-W436F-mCherry.
	Figure 2. NavBeta effect on Kv1.3 gating currents in rodents and human.(A) Amino acid sequence alignment of human NavBeta1 (hNavBeta1) and rat NavBeta1 (rNavBeta1). Residues that differ between them are highlighted in red. Transmembrane segment is indicated with an orange rectangle. W172 is indicated in purple circle (See also Fig. 5). (B,C) Q-V relationship (B) and time constants of on-gating currents as a function of voltage (Tau-V) (C) for hKv1.3-del-W436F alone (black circles, n = 8), and co-injected with rNavBeta1 (red circles, n = 5) or with hNavBeta1 (blue circles, n = 5). Red stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del-W436F alone (black circles) and with rNavBeta1 (red circles) at voltages ranging from −50 mV to +10 mV in Q-V (B), and at voltages ranging from −15 mV to +40 mV for Tau-V (C). Blue stars (*) indicate statistical significance (p-value < 0.05) between hKv1.3-del-W436F alone (black circles) and with hNavBeta1 (blue circles) at voltages ranging from −45 mV to −10 mV for Q-V (B), and at voltages ranging from −15 mV to +40 mV for Tau-V (C). (D,E) Q-V relationship (D) and Tau-V (E) for mouse Kv1.3 (mKv1.3)-W389F alone (black triangles, n = 4) and co-injected with rNavBeta1 (red triangles, n = 4). Red stars (*) indicate statistical significance (p-value < 0.05) of the difference between mKv1.3-W389F alone (black) and with rNavBeta1 (red) for voltages ranging from −55 mV to −10 mV in Q-V (D), and at voltages ranging from 0 mV to +40 mV in Tau-V (E). The pulse protocol for gating current measurements is depicted in the upper right corner in C). Error bars indicate SEM.
	Figure 3. Gating currents from hKv1.3-del-W436F and conductive hKv1.3-del with/without rNavBeta1.(A–D) Representative gating currents from hKv1.3-del-W436F alone (A), co-injected with rNavBeta1 (B), from conductive hKv1.3-del alone while blocked by Shk Tx (C) and co-injected with rNavBeta1 (D). (E,F) The Q-V relationship (E) and Tau-V relationship (F) for hKv1.3-del-W436F alone (filled black circles, n = 8) and co-injected with rNavBeta1 (filled red circles, n = 5), and for conductive hKv1.3-del blocked by ShkTx (opened black circles, n = 4) and co-injected with rNavBeta1 (opened red circles, n = 6). Error bars indicate SEM. Red star (*) indicates statistical significance (p-value < 0.05) of the difference between conductive hKv1.3-del blocked by ShkTx (opened black circles) and with rNavBeta1 (opened red circles) from −115 mV to +15 mV in Q-V (E), and from −10 mV to +40 mV in Tau-V (F).
	Figure 4. The effect of chimeras of rNavBeta1 and P0 on hKv1.3 gating current.(A) Amino acid sequence alignment between rNavBeta1 and myelin protein zero (P0), a protein with high homology to rNavBeta1. Transmembrane segment is highlighted by orange rectangle. The sequence for NavBeta1-P0 chimera is indicated in green and that for P0-NavBeta1 in purple. (B,C) Representative gating currents from hKv1.3-del-W436F co-injected with P0-NavBeta1 (B) or with NavBeta1-P0 (C). (D,E) The Q-V relationship (D) and Tau-V relationship (E) for hKv1.3-del-W436F alone (black circles, n = 8), co-injected with rNavBeta1 (red circles, n = 5), co-injected with P0-NavBeta1 (purple squares, n = 5) or co-injected with NavBeta1-P0 (green squares, n = 3). Error bars indicate SEM. Green stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del-W436F alone (black circles) and with NavBeta1-P0 (green squares) from −95 mV to −10 mV in Q-V (D), and at −15, −10, +5, +20 and +40 mV in Tau-V (E). Purple stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del-W436F alone (black circles) and with P0-NavBeta1 (purple squares) from −70 mV to +20 mV in Q-V (D), and from −40 mV to +40 mV except at −20 mV in Tau-V (E).
	Figure 5. W172 in NavBeta1 transmembrane segment is part of an interaction interface with hKv1.3.(A,B) Representative gating currents from hKv1.3-del-W436F co-injected with P0-NavBeta1-W172A (A) or with NavBeta1-W172A (B). (C,D) The Q-V relationship (C) and Tau-V curve (D) for hKv1.3-del-W436F alone (black circles, n = 8), co-injected with rNavBeta1 (red circles, n = 5), co-injected with P0-NavBeta1 (purple squares, n = 5), co-injected with P0-NavBeta1-W172A (orange squares, n = 3) or co-injected with NavBeta1-W172A (blue circles, n = 3). Error bars indicate SEM. Orange stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del-W436F with P0-NavBeta1 (purple squares) and with P0-NavBeta1-W172A (orange squares) from −30 mV to +5 mV in Q-V (C), and from −40 mV to +40 mV except at −10 mV in Tau-V (D). Blue stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del-W436F with rNavBeta1 (red circles) and with NavBeta1-W172A (blue circles) from −45 mV to −25 mV except at −40 mV in Q-V (C), and from −15 mV to +40 mV in Tau-V (D).
	Figure 6. NavBeta1 effect on hKv1.3-del ionic currents.(A) Representative ionic currents from hKv1.3-del alone (black, n = 5), co-injected with rNavBeta1 (red, n = 4) or co-injected with hNavBeta1 (blue, n = 4). Pulse protocol is shown at the top of the panel. (B) Normalized conductance as a function of voltage (G-V) for hKv1.3-del alone (black), co-injected with rNavBeta1 (red) or with hNavBeta1 (blue). (C) Time constants of activation as a function of voltage for hKv1.3-del alone (black), co-injected with rNavBeta1 (red) or co-injected with hNavBeta1 (blue). Time constants were obtained as weighted average values from the fit of the rising face of the currents (highlighted in yellow in A) to two exponential functions. Blue star (*) indicates statistical significance (p-value < 0.05) of the difference between hKv1.3-del alone (black) and with hNavBeta1 (blue) from +10 mV to +80 mV except at +20 mV. (D) Deactivation time constants from the tail currents at –80 mV (highlighted in yellow at the end of the pulse in A) for hKv1.3-del alone (black), co-injected with rNavBeta1 (red) or co-injected with hNavBeta1 (blue). Time constants were obtained as weighted average values from the fit to two exponentials. Stars (*) indicate statistical significance (p-value < 0.05) of the difference between hKv1.3-del alone (black) and with rNavBeta1 (red) in red, and with hNavBeta1 (blue) in blue, from −20 mV to +80 mV. (E) Comparison between G-V and Q-V between hKv1.3 alone and when co-injected with rNavBeta1. To reach 50% of the maximum conductance (Gmax), hKv1.3 with rNavBeta1 requires less charge movement than hKv1.3 alone, indicating rNavBeta1 may strengthen the coupling between VSD movement and pore opening.

Attali, Cloning, functional expression, and regulation of two K+ channels in human T lymphocytes. 1992, Pubmed, Xenbase

Attali, Cloning, functional expression, and regulation of two K+ channels in human T lymphocytes. 1992, Pubmed , Xenbase
Azam, Targeting effector memory T cells with the small molecule Kv1.3 blocker PAP-1 suppresses allergic contact dermatitis. 2007, Pubmed
Beeton, Kv1.3 channels are a therapeutic target for T cell-mediated autoimmune diseases. 2006, Pubmed
Beeton, Selective blocking of voltage-gated K+ channels improves experimental autoimmune encephalomyelitis and inhibits T cell activation. 2001, Pubmed
Beeton, Selective blockade of T lymphocyte K(+) channels ameliorates experimental autoimmune encephalomyelitis, a model for multiple sclerosis. 2001, Pubmed
Black, Noncanonical roles of voltage-gated sodium channels. 2013, Pubmed
Brackenbury, Functional reciprocity between Na+ channel Nav1.6 and beta1 subunits in the coordinated regulation of excitability and neurite outgrowth. 2010, Pubmed
Brackenbury, Abnormal neuronal patterning occurs during early postnatal brain development of Scn1b-null mice and precedes hyperexcitability. 2013, Pubmed
Brackenbury, Voltage-gated Na+ channel beta1 subunit-mediated neurite outgrowth requires Fyn kinase and contributes to postnatal CNS development in vivo. 2008, Pubmed
Cahalan, A voltage-gated potassium channel in human T lymphocytes. 1985, Pubmed
Calhoun, The role of non-pore-forming β subunits in physiology and pathophysiology of voltage-gated sodium channels. 2014, Pubmed
Candenas, Molecular diversity of voltage-gated sodium channel alpha and beta subunit mRNAs in human tissues. 2006, Pubmed
Cha, Structural implications of fluorescence quenching in the Shaker K+ channel. 1998, Pubmed
Coleman, Subunit composition of Kv1 channels in human CNS. 1999, Pubmed
Dang, Native chemical ligation at Asx-Cys, Glx-Cys: chemical synthesis and high-resolution X-ray structure of ShK toxin by racemic protein crystallography. 2013, Pubmed
Davis, Sodium channel beta1 subunits promote neurite outgrowth in cerebellar granule neurons. 2004, Pubmed
DeCoursey, Voltage-gated K+ channels in human T lymphocytes: a role in mitogenesis? , Pubmed
Deschênes, Modulation of Kv4.3 current by accessory subunits. 2002, Pubmed
Deschênes, Post-transcriptional gene silencing of KChIP2 and Navbeta1 in neonatal rat cardiac myocytes reveals a functional association between Na and Ito currents. 2008, Pubmed
Douglass, Characterization and functional expression of a rat genomic DNA clone encoding a lymphocyte potassium channel. 1990, Pubmed , Xenbase
Fukushima, Potassium current in clonal cytotoxic T lymphocytes from the mouse. 1984, Pubmed
Ghanshani, Up-regulation of the IKCa1 potassium channel during T-cell activation. Molecular mechanism and functional consequences. 2000, Pubmed
Gilhar, The beneficial effect of blocking Kv1.3 in the psoriasiform SCID mouse model. 2011, Pubmed
Grissmer, Expression and chromosomal localization of a lymphocyte K+ channel gene. 1990, Pubmed , Xenbase
Grissmer, The Shaw-related potassium channel gene, Kv3.1, on human chromosome 11, encodes the type l K+ channel in T cells. 1992, Pubmed , Xenbase
Hondares, Enhanced activation of an amino-terminally truncated isoform of the voltage-gated proton channel HVCN1 enriched in malignant B cells. 2014, Pubmed
Hyodo, Voltage-gated potassium channel Kv1.3 blocker as a potential treatment for rat anti-glomerular basement membrane glomerulonephritis. 2010, Pubmed
Ishida, Lack of voltage sensitive potassium channels and generation of membrane potential by sodium potassium ATPase in murine T lymphocytes. 1993, Pubmed
Isom, Primary structure and functional expression of the beta 1 subunit of the rat brain sodium channel. 1992, Pubmed , Xenbase
Koch, Complex subunit assembly of neuronal voltage-gated K+ channels. Basis for high-affinity toxin interactions and pharmacology. 1997, Pubmed
Koo, Blockade of the voltage-gated potassium channel Kv1.3 inhibits immune responses in vivo. 1997, Pubmed
Kundu-Raychaudhuri, Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis--xenograft model. 2014, Pubmed
Labro, Molecular mechanism for depolarization-induced modulation of Kv channel closure. 2012, Pubmed , Xenbase
Lemke, Isolation and sequence of a cDNA encoding the major structural protein of peripheral myelin. 1985, Pubmed
Lioudyno, Shaker-related potassium channels in the central medial nucleus of the thalamus are important molecular targets for arousal suppression by volatile general anesthetics. 2013, Pubmed
Litan, Cancer as a channelopathy: ion channels and pumps in tumor development and progression. 2015, Pubmed
Liu, Modulation of Kv channel expression and function by TCR and costimulatory signals during peripheral CD4(+) lymphocyte differentiation. 2002, Pubmed
Marionneau, The sodium channel accessory subunit Navβ1 regulates neuronal excitability through modulation of repolarizing voltage-gated K⁺ channels. 2012, Pubmed
Matheu, Imaging of effector memory T cells during a delayed-type hypersensitivity reaction and suppression by Kv1.3 channel block. 2008, Pubmed
Matteson, K channels in T lymphocytes: a patch clamp study using monoclonal antibody adhesion. , Pubmed
McCormick, The extracellular domain of the beta1 subunit is both necessary and sufficient for beta1-like modulation of sodium channel gating. 1999, Pubmed , Xenbase
Nguyen, Modulation of voltage-gated K+ channels by the sodium channel β1 subunit. 2012, Pubmed , Xenbase
O'Malley, Sodium channel β subunits: emerging targets in channelopathies. 2015, Pubmed
Patton, The adult rat brain beta 1 subunit modifies activation and inactivation gating of multiple sodium channel alpha subunits. 1994, Pubmed , Xenbase
Pereira, Pharmacokinetics, toxicity, and functional studies of the selective Kv1.3 channel blocker 5-(4-phenoxybutoxy)psoralen in rhesus macaques. 2007, Pubmed
Perozo, Gating currents from a nonconducting mutant reveal open-closed conformations in Shaker K+ channels. 1993, Pubmed
Rus, The voltage-gated potassium channel Kv1.3 is highly expressed on inflammatory infiltrates in multiple sclerosis brain. 2005, Pubmed
Stefani, Cut-open oocyte voltage-clamp technique. 1998, Pubmed , Xenbase
Tarcha, Durable pharmacological responses from the peptide ShK-186, a specific Kv1.3 channel inhibitor that suppresses T cell mediators of autoimmune disease. 2012, Pubmed
Upadhyay, Selective Kv1.3 channel blocker as therapeutic for obesity and insulin resistance. 2013, Pubmed
Vicente, Association of Kv1.5 and Kv1.3 contributes to the major voltage-dependent K+ channel in macrophages. 2006, Pubmed , Xenbase
Wulff, The voltage-gated Kv1.3 K(+) channel in effector memory T cells as new target for MS. 2003, Pubmed