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Science 20 December 1996:
Vol. 274. no. 5295, pp. 2089 - 2091
DOI: 10.1126/science.274.5295.2089
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Reports

Association of Src Tyrosine Kinase with a Human Potassium Channel Mediated by SH3 Domain

Todd C. Holmes, Debra A. Fadool, Ruibao Ren, Irwin B. Levitan *

The human Kv1.5 potassium channel (hKv1.5) contains proline-rich sequences identical to those that bind to Src homology 3 (SH3) domains. Direct association of the Src tyrosine kinase with cloned hKv1.5 and native hKv1.5 in human myocardium was observed. This interaction was mediated by the proline-rich motif of hKv1.5 and the SH3 domain of Src. Furthermore, hKv1.5 was tyrosine phosphorylated, and the channel current was suppressed, in cells coexpressing v-Src. These results provide direct biochemical evidence for a signaling complex composed of a potassium channel and a protein tyrosine kinase.

T. C. Holmes, D. A. Fadool, I. B. Levitan, Department of Biochemistry and Volen Center for Complex Systems, Brandeis University, Waltham, MA 02254, USA.
R. Ren, Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02254, USA.
*   To whom correspondence should be addressed.

Potassium channels are important for such cellular electrical properties as resting potential, excitability, and the repolarization of the action potential. Thus, modulation of these channels can profoundly affect physiological processes including neuronal integration, vesicle secretion, and muscle contraction. The modulation of potassium channel activity by serine-threonine kinases has been studied extensively (1). The recently discovered PYK2 tyrosine kinase (2), as well as endogenous tyrosine kinases in human embryonic kidney (HEK) 293 cells (3), can also phosphorylate and suppress the activity of potassium channels. In spite of emerging evidence concerning the functional effects of tyrosine phosphorylation of potassium channels, there is no information available about the mechanisms of targeting and association of these channels with tyrosine kinases. However, the existence of signaling complexes consisting of ion channels and closely associated protein kinases and phosphatases has been inferred from biochemical and functional electrophysiological studies (4).

Specific protein-protein interactions between signaling proteins are mediated by modular binding domains (5). Among the first of these to be characterized was a conserved sequence found in the Src tyrosine kinase, known as the Src homology 3 (SH3) domain. SH3 domains bind to proline-rich regions in partner proteins. We examined the sequences of mammalian voltage-dependent potassium channels, and noted that several species isoforms of Kv1.5--including those from human (hKv1.5), dog, and rabbit (6)--contain one to two copies of the preferred Src SH3 domain binding motif RPLPXXP (7, 8). In particular, hKv1.5 contains two repeats of the sequence RPLPPLP between amino acid residues 65 and 82 of the channel protein (6, 8, 9). To determine whether hKv1.5 and Src are associated in vivo, we coexpressed the channel and kinase in HEK 293 cells and tested for their interaction by immunoprecipitation followed by protein immunoblotting with specific antibodies to hKv1.5 and Src (10).

When hKv1.5 and associated proteins were immunoprecipitated from cell lysates with a specific antibody, Src was co-precipitated (Fig. 1A). Similarly, when Src and associated proteins were immunoprecipitated from HEK 293 cell lysates, hKv1.5 co-precipitated with endogenous and coexpressed Src (Fig. 1A). Expression of hKv1.5 protein was not altered by v-Src coexpression, as verified by protein immunoblot analysis of cell lysates with antibodies directed against tagged (Fig. 1A) and native (Fig. 2A) sequences of the channel. Furthermore, immunoblot (Fig. 1A) or protein silver stain (Fig. 3A) analysis of immunoprecipitates demonstrated that the efficiency of immunoprecipitation of hKv1.5 was not affected by v-Src coexpression. Enzymatic activity of Src also co-precipitated with hKv1.5, as detected by an in vitro kinase assay with hKv1.5 immunoprecipitates and an Src-specific substrate (11) (Fig. 1B).

Fig. 1. Co-immunoprecipitation of hKv1.5 and Src. (A) HEK 293 cells were transfected with CMV vector with no insert (control), vector encoding v-Src, vector encoding hKv1.5, or two separate vectors, one encoding hKv1.5 and the other encoding v-Src (10). Expression of hKv1.5 was measured on protein immunoblots of cell lysates probed with anti-tag-hKv1.5 (top panel). Densitometry of autoradiograms was used to quantitate channel expression. For each experiment, the density (expression level) of hKv1.5 transfected alone was set to 1. When Src was cotransfected, the relative density of hKv1.5 was 0.98 ± 0.05 (mean ± SEM; n = 9; not significant by t test). Immunoblot analysis of anti-tag-hKv1.5 immunoprecipitates (IP) with anti-tag-hKv1.5 probe (second panel) confirmed that the efficiency of immunoprecipitation of hKv1.5 (density set to 1) was not affected by coexpression of v-Src (relative density 1.06 ± 0.10; n = 8; not significant by t test). Src that co-immunoprecipitated with hKv1.5 was detected by probing anti-tag-hKv1.5 IP with anti-Src (third panel). The hKv1.5 that co-immunoprecipitated with endogenous c-Src and expressed v-Src was detected by probing anti-Src IP with anti-tag-hKv1.5 (bottom panel). (B) HEK 293 cells were transfected as in (A). Anti-tag-hKv1.5 IP were assayed for Src activity (21) by incubation with or without the Src substrate fusion protein G10A. The reaction products were separated on protein immunoblots that were probed with antibody 4G10 to phosphotyrosine (n = 4). (C) Native Src that co-immunoprecipitated with native Kv1.5, or was immunoprecipitated directly with anti-Src, was detected by probing IP prepared from human myocardium tissue lysates (separated on immunoblots) with anti-Src (n = 4). IP with an antiserum (3) against Kv1.3, another potassium channel, was used as an additional control. [View Larger Version of this Image (35K GIF file)]

Fig. 2. Domains that mediate binding of hKv1.5 to Src. (A) HEK 293 cells were transfected with CMV vector with no insert (control), vector encoding v-Src, vector encoding hKv1.5 or rKv1.5, or two separate vectors, one encoding hKv1.5 or rKv1.5 and the other encoding v-Src (10, 15). Expression of hKv1.5 and rKv1.5 was detected in cell lysates by protein immunoblotting with anti-Kv1.5 (19), which recognizes both channels (top panel). Because this antibody recognizes rKv1.5 much better than hKv1.5, the two parts of this panel were exposed for different times. The apparent doublet in the rKv1.5 lanes may represent phosphorylated or glycosylated or both types of isoforms of the channel protein (3). Anti-Src IP, separated on immunoblots, were probed with anti-Kv1.5 (bottom panel) (n = 4). (B) HEK 293 cells were transfected with CMV vectors coding for vector with no insert (control) or hKv1.5. Expression of hKv1.5 was confirmed by immunoblotting the cell lysates with anti-tag-hKv1.5 (top panel). Cell lysates were incubated with GST alone or GST fusion protein containing the Src SH3 domain (GST-Src-SH3) (22), with or without fusion protein preabsorption with a peptide corresponding to the proline-rich sequence comprising amino acids 62 through 83 of hKv1.5 (peptide hKv1.562-83) (15). Proteins bound to GST fusion proteins were separated by SDS-PAGE, and immunoblots were probed with anti-tag-hKv1.5 (bottom panel) (n = 4). (C) Far western blots were prepared with anti-Kv1.5 IP from cells transfected with hKv1.5 or rKv1.5. The blots were probed with biotinylated GST-Src-SH3 (1 µg/ml) (22) (top panel; the arrow indicates position of the hKv1.5 band) or biotinylated GST-Src-SH3 preabsorbed with peptide hKv1.562-83 (bottom panel). The blots were then incubated with avidin-horseradish peroxidase, and bound fusion protein was visualized by ECL (n = 4). [View Larger Version of this Image (34K GIF file)]


Fig. 3. Tyrosine phosphorylation of hKv1.5 and suppression of channel current by coexpression with v-Src. (A) HEK 293 cells were transfected with CMV vectors as indicated: vector with no insert (control); v-Src; hKv1.5; or one vector encoding hKv1.5 and another encoding v-Src. Cells were lysed, and proteins were immunoprecipitated with anti-tag-hKv1.5. IP were separated by SDS-PAGE, and protein was detected by silver stain (20) (top panel). Immunoblots (bottom panel) were probed with antibody 4G10 to phosphotyrosine (n = 4). (B) HEK 293 cells were transfected with a CMV vector encoding hKv1.5 or one vector encoding hKv1.5 and another encoding v-Src. Macroscopic currents evoked by a series of depolarizing voltage pulses were recorded from cell-attached membrane patches (3, 16) 2 days after transfection. The peak current at +40 mV was 592 ± 163 pA (mean ± SEM; n = 8) in patches from cells expressing hKv1.5 alone, and 27 ± 15 pA (n = 9) in patches from cells coexpressing v-Src (significantly different, P <= 0.02, t-test). [View Larger Versions of these Images (41K GIF file)]

The association between hKv1.5 and Src was also observed in human tissue. Native Src was detected in immunoprecipitates, prepared with a Kv1.5 antiserum, from human myocardium ventricle tissue lysates (Fig. 1C). The native Src that co-immunoprecipitated with native Kv1.5 co-migrated on protein immunoblots with native Src, immunoprecipitated directly with a polyclonal anti-Src antibody (Fig. 1C). Thus association of hKv1.5 and Src occurs under physiological conditions, and does not depend on expression in a heterologous system. This association may contribute to the co-localization of Kv1.5 and Src in cellular adhesion zones in myocardium (12). Although the stoichiometry of the association between hKv1.5 and Src is not known, only a fraction of the total myocardial Src co-immunoprecipitated with hKv1.5 (Fig. 1C), consistent with the fact that Src phosphorylates other substrates.

There are specific sequence requirements for the association of hKv1.5 and Src. For example the NH2-terminal region of the rat Kv1.5 (rKv1.5) channel also contains a proline-rich motif (9, 13), but this sequence (RPLPPMA) (8) does not appear to be favorable for binding to the Src SH3 domain, as shown by the absence of selection of this sequence with phage display libraries (7). In contrast to hKv1.5, rKv1.5 failed to co-immunoprecipitate with Src (Fig. 2A). Thus, the association between channel and Src is detected only for the hKv1.5 channel isoform, possibly because its proline-rich binding motif is preferred by the Src SH3 domain. In addition phospholipase C-gamma and the p85 regulatory subunit of phosphatidylinositol 3-kinase, which contain SH3 domains with different binding sequence requirements than that of Src (7), do not co-immunoprecipitate with hKv1.5 (14).

We tested hKv1.5 binding to the Src SH3 domain itself expressed as a fusion protein with glutathione-S-transferase (GST) (15). Cell lysates prepared from vector control and hKv1.5 transfected cells were incubated with a GST fusion protein containing the Src SH3 domain (GST-Src-SH3) or no insert (GST). The hKv1.5 protein was effectively precipitated by GST-Src-SH3, but not by GST (Fig. 2B). The specificity of this interaction was tested by preabsorption of the fusion proteins with a peptide containing the sequence of the proline-rich region of hKv1.5 (peptide hKv1.562-83). Binding of hKv1.5 to GST-Src-SH3 was attenuated by preabsorption of the fusion protein with peptide Kv1.562-83 (Fig. 2B). The direct binding of the Src SH3 domain to hKv1.5 was demonstrated in a filter binding assay (far Western blot). GST-Src-SH3 bound to hKv1.5 on the filter, whereas no binding was detected with rKv1.5 (Fig. 2C). The role of the proline-rich motif in the channel in the binding of GST-Src-SH3 to hKv1.5 was demonstrated further by the absence of filter binding after preabsorption of the GST-Src-SH3 with peptide hKv1.562-83 (Fig. 2C).

The hKv1.5 protein was tyrosine phosphorylated when it was coexpressed with v-Src (Fig. 3A). To determine whether coexpression of v-Src influenced channel activity, we measured hKv1.5 macroscopic currents in cell-attached membrane patches, with and without v-Src coexpression (3, 16). Current through hKv1.5 channels was suppressed when the channel was coexpressed with v-Src (Fig. 3B), even though channel protein expression was not altered (Fig. 3A; see also Figs. 1A and 2A). We do not yet know whether the suppression of hKv1.5 current (Fig. 3B) results from tyrosine phosphorylation of the channel protein (Fig. 3A), or whether direct binding of hKv1.5 to Src is required for either of these phenomena.

Many mammalian ion channels, including channels that are known to be modulated by tyrosine kinases, have proline-rich sequences that may bind to SH3 domains (9). Furthermore, signaling complexes containing multiple protein kinases, or ion channels together with scaffolding and regulatory proteins, may be common in cells (4, 17). Potential consequences of a closely associated channel-kinase signaling complex include increased specificity of signaling pathways, faster coupling, and a higher probability of channel phosphorylation after kinase activation.

REFERENCES AND NOTES

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  15. HEK 293 cells were transfected as described (3, 10). The CMV vector encoding rKv1.5 was provided by J. Trimmer. For fusion protein precipitation experiments, cell lysates were incubated overnight at 4°C with GST fusion protein (1.0 µg/ml of cell lysate) (22). The GST fusion proteins and bound proteins were separated from unbound protein by incubation with glutathione-agarose beads. Where indicated, GST fusion proteins were preabsorbed by incubating with 100 times greater concentration of peptide NH2-SGVRPLPPLPDPGVRPLPPLPE-COOH (peptide hKv1.562-83; Bio-Synthesis, Lewisville, TX) (8). The proteins bound to the washed agarose beads were separated by SDS-PAGE, and immunoblots were probed with anti-tag-hKv1.5.
  16. Patch-clamp recordings of macroscopic currents were made in the cell-attached patch configuration (3). Patches were held at -90 mV and stepped to depolarizing potentials in 5-mV increments to 0 mV. The pulse duration was 400 ms and the interpulse interval was 10 s.
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  23. We thank R. Huganir, L. Philipson, and J. Trimmer for antibodies and cDNA constructs; M. Mendelsohn for samples of human myocardial tissue; and C. Miller for comments on the manuscript. Supported by grants to I.B.L. and R.R. from NIH. T.C.H. and D.A.F. were supported by National Research Service Awards.

26 June 1996; accepted 22 October 1996


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B. Soliven, L. Ma, H. Bae, B. Attali, A. Sobko, and T. Iwase (2003)
Am J Physiol Cell Physiol 284, C85-C93
   Abstract »    Full Text »    PDF »
Zinc Mediates Assembly of the T1 Domain of the Voltage-gated K Channel 4.2.
A. W. Jahng, C. Strang, D. Kaiser, T. Pollard, P. Pfaffinger, and S. Choe (2002)
J. Biol. Chem. 277, 47885-47890
   Abstract »    Full Text »    PDF »
Coupling of c-Src to large conductance voltage- and Ca2+-activated K+ channels as a new mechanism of agonist-induced vasoconstriction.
A. Alioua, A. Mahajan, K. Nishimaru, M. M. Zarei, E. Stefani, and L. Toro (2002)
PNAS 99, 14560-14565
   Abstract »    Full Text »    PDF »
Tyrosine Phosphorylation of Kv1.2 Modulates Its Interaction with the Actin-binding Protein Cortactin.
D. Hattan, E. Nesti, T. G. Cachero, and A. D. Morielli (2002)
J. Biol. Chem. 277, 38596-38606
   Abstract »    Full Text »    PDF »
Phosphorylation-Dependent and Phosphorylation-Independent Modes of Modulation of Shaker Family Voltage-Gated Potassium Channels by Src Family Protein Tyrosine Kinases.
M. N. Nitabach, D. A. Llamas, I. J. Thompson, K. A. Collins, and T. C. Holmes (2002)
J. Neurosci. 22, 7913-7922
   Abstract »    Full Text »    PDF »
Calmodulin Is an Auxiliary Subunit of KCNQ2/3 Potassium Channels.
H. Wen and I. B. Levitan (2002)
J. Neurosci. 22, 7991-8001
   Abstract »    Full Text »    PDF »
N-terminal Tyrosine Residues within the Potassium Channel Kir3 Modulate GTPase Activity of Galpha i.
D. L. Ippolito, P. A. Temkin, S. L. Rogalski, and C. Chavkin (2002)
J. Biol. Chem. 277, 32692-32696
   Abstract »    Full Text »    PDF »
Amino-terminal Determinants of U-type Inactivation of Voltage-gated K+ Channels.
H. T. Kurata, G. S. Soon, J. R. Eldstrom, G. W. K. Lu, D. F. Steele, and D. Fedida (2002)
J. Biol. Chem. 277, 29045-29053
   Abstract »    Full Text »    PDF »
Neurotrophin modulation of voltage-gated potassium channels in rat through TrkB receptors is time and sensory experience dependent.
K Tucker and D. Fadool (2002)
J. Physiol. 542, 413-429
   Abstract »    Full Text »    PDF »
Effects of Protein Kinase C on Delayed Rectifier K+ Channel Regulation by Tyrosine Kinase in Rat Retinal Pigment Epithelial Cells.
O. Strauss, R. Rosenthal, D. Dey, J. Beninde, G. Wollmann, H. Thieme, and M. Wiederholt (2002)
Invest. Ophthalmol. Vis. Sci. 43, 1645-1654
   Abstract »    Full Text »    PDF »
Regulation of an ERG K+ Current by Src Tyrosine Kinase.
F. S. Cayabyab and L. C. Schlichter (2002)
J. Biol. Chem. 277, 13673-13681
   Abstract »    Full Text »    PDF »
Two Adaptor Proteins Differentially Modulate the Phosphorylation and Biophysics of Kv1.3 Ion Channel by Src Kinase.
K. K. Cook and D. A. Fadool (2002)
J. Biol. Chem. 277, 13268-13280
   Abstract »    Full Text »    PDF »
Regulated Cationic Channel Function in Xenopus Oocytes Expressing Drosophila Big Brain.
G. M. Yanochko and A. J. Yool (2002)
J. Neurosci. 22, 2530-2540
   Abstract »    Full Text »    PDF »
Signal Transduction of Physiological Concentrations of Vasopressin in A7r5 Vascular Smooth Muscle Cells. A ROLE FOR PYK2 AND TYROSINE PHOSPHORYLATION OF K+ CHANNELS IN THE STIMULATION OF Ca2+ SPIKING.
K. L. Byron and P. A. Lucchesi (2002)
J. Biol. Chem. 277, 7298-7307
   Abstract »    Full Text »    PDF »
Modulation of Kv1.5 Currents by Protein Kinase A, Tyrosine Kinase, and Protein Tyrosine Phosphatase Requires an Intact Cytoskeleton.
H. S. Mason, M. J. Latten, L. D. Godoy, B. Horowitz, and J. L. Kenyon (2002)
Mol. Pharmacol. 61, 285-293
   Abstract »    Full Text »    PDF »
Protein Kinase Modulation of a Neuronal Cation Channel Requires Protein-Protein Interactions Mediated by an Src homology 3 Domain.
N. S. Magoski, G. F. Wilson, and L. K. Kaczmarek (2002)
J. Neurosci. 22, 1-9
   Abstract »    Full Text »    PDF »
Regulation of ion channels by protein tyrosine phosphorylation.
M. J. Davis, X. Wu, T. R. Nurkiewicz, J. Kawasaki, P. Gui, M. A. Hill, and E. Wilson (2001)
Am J Physiol Heart Circ Physiol 281, H1835-H1862
   Abstract »    Full Text »    PDF »
Unzipping Ion Channels.
S. N. MacFarlane and I. B. Levitan (2001)
Sci. STKE 2001, pe1
   Abstract »    Full Text »    PDF »
Modulation of Excitability in Aplysia Tail Sensory Neurons by Tyrosine Kinases.
A. L. Purcell and T. J. Carew (2001)
J Neurophysiol 85, 2398-2411
   Abstract »    Full Text »    PDF »
Adenosine 5'-Triphosphate: a P2-Purinergic Agonist in the Myocardium.
G. Vassort (2001)
Physiol Rev 81, 767-806
   Abstract »    Full Text »    PDF »
Tyrosine Decaging Leads to Substantial Membrane Trafficking during Modulation of an Inward Rectifier Potassium Channel.
Y. Tong, G. S. Brandt, M. Li, G. Shapovalov, E. Slimko, A. Karschin, D. A. Dougherty, and H. A. Lester (2001)
J. Gen. Physiol. 117, 103-118
   Abstract »    Full Text »    PDF »
A mechanism for combinatorial regulation of electrical activity: Potassium channel subunits capable of functioning as Src homology 3-dependent adaptors.
M. N. Nitabach, D. A. Llamas, R. C. Araneda, J. L. Intile, I. J. Thompson, Y. I. Zhou, and T. C. Holmes (2001)
PNAS
   Abstract »    Full Text »
Cell cycle-related changes in transient K+ current density in the GH3 pituitary cell line.
A. Czarnecki, S. Vaur, L. Dufy-Barbe, B. Dufy, and L. Bresson-Bepoldin (2000)
Am J Physiol Cell Physiol 279, C1819-C1828
   Abstract »    Full Text »    PDF »
Opening and Closing of KcnkO Potassium Leak Channels Is Tightly Regulated.
N. Zilberberg, N. Ilan, R. Gonzalez-Colaso, and S. A.N. Goldstein (2000)
J. Gen. Physiol. 116, 721-734
   Abstract »    Full Text »    PDF »
Physiology of apoptosis.
E. Gulbins, A. Jekle, K. Ferlinz, H. Grassme, and F. Lang (2000)
Am J Physiol Renal Physiol 279, F605-F615
   Abstract »    Full Text »    PDF »
Modulation of Kv1.5 Currents by Src Tyrosine Phosphorylation: Potential Role in the Differentiation of Astrocytes.
S. N. MacFarlane and H. Sontheimer (2000)
J. Neurosci. 20, 5245-5253
   Abstract »    Full Text »    PDF »
Interactions of Cyclic Nucleotide-Gated Channel Subunits and Protein Tyrosine Kinase Probed with Genistein.
E. Molokanova, A. Savchenko, and R. H. Kramer (2000)
J. Gen. Physiol. 115, 685-696
   Abstract »    Full Text »    PDF »
Interaction of the N-Methyl-D-Aspartic Acid Receptor NR2D Subunit with the c-Abl Tyrosine Kinase.
R. T. Glover, M. Angiolieri, S. Kelly, D. T. Monaghan, J. Y. J. Wang, T. E. Smithgall, and A. L. Buller (2000)
J. Biol. Chem. 275, 12725-12729
   Abstract »    Full Text »    PDF »
RPTP{micro} and protein tyrosine phosphorylation regulate K+ channel mRNA expression in adult cardiac myocytes.
K. M. Hershman and E. S. Levitan (2000)
Am J Physiol Cell Physiol 278, C397-C403
   Abstract »    Full Text »    PDF »
Phosphorylation Is Required for Alteration of Kv1.5 K+ Channel Function by the Kvbeta 1.3 Subunit.
Y.-G. Kwak, R. A. Navarro-Polanco, T. Grobaski, D. J. Gallagher, and M. M. Tamkun (1999)
J. Biol. Chem. 274, 25355-25361
   Abstract »    Full Text »    PDF »
Tyrosine kinases modulate K+ channel gating in mouse Schwann cells.
A. Peretz, A. Sobko, and B. Attali (1999)
J. Physiol. 519, 373-384
   Abstract »    Full Text »    PDF »
Activity-Dependent Modulation of Rod Photoreceptor Cyclic Nucleotide-Gated Channels Mediated by Phosphorylation of a Specific Tyrosine Residue.
E. Molokanova, F. Maddox, C. W. Luetje, and R. H. Kramer (1999)
J. Neurosci. 19, 4786-4795
   Abstract »    Full Text »    PDF »
HIV-1 Nef Expression Inhibits the Activity of a Ca2+-Dependent K+ Channel Involved in the Control of the Resting Potential in CEM Lymphocytes.
O. Zegarra-Moran, A. Rasola, M. Rugolo, A. M. Porcelli, B. Rossi, and L. J. V. Galietta (1999)
J. Immunol. 162, 5359-5366
   Abstract »    Full Text »    PDF »
Structure and function of cardiac potassium channels.
D. J Snyders (1999)
Cardiovasc Res 42, 377-390
   Abstract »    Full Text »    PDF »
Growth factor-mediated Fyn signaling regulates alpha -amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor expression in rodent neocortical neurons.
M. Narisawa-Saito, A. J. Silva, T. Yamaguchi, T. Hayashi, T. Yamamoto, and H. Nawa (1999)
PNAS 96, 2461-2466
   Abstract »    Full Text »    PDF »
An Ultraviolet-activated K+ Channel Mediates Apoptosis Of Myeloblastic Leukemia Cells.
L. Wang, D. Xu, W. Dai, and L. Lu (1999)
J. Biol. Chem. 274, 3678-3685
   Abstract »    Full Text »    PDF »
The fyn art of N-methyl-D-aspartate receptor phosphorylation.
C. Sala and M. Sheng (1999)
PNAS 96, 335-337
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Noncatalytic Inhibition of Cyclic Nucleotide-gated Channels by Tyrosine Kinase Induced by Genistein.
E. Molokanova, A. Savchenko, and R. H. Kramer (1999)
J. Gen. Physiol. 113, 45-56
   Abstract »    Full Text »    PDF »
Tyrosine kinase involvement in apamin-sensitive inhibitory responses of rat distal colon.
T. Takeuchi, M. Kishi, N. Hirayama, M. Yamaji, T. Ishii, H. Nishio, F. Hata, and T. Takewaki (1999)
J. Physiol. 514, 177-188
   Abstract »    Full Text »    PDF »
Acute Suppression of Inwardly Rectifying Kir2.1 Channels by Direct Tyrosine Kinase Phosphorylation.
E. Wischmeyer, F. Doring, and A. Karschin (1998)
J. Biol. Chem. 273, 34063-34068
   Abstract »    Full Text »    PDF »
Differential regulation of potassium currents by FGF-1 and FGF-2 in embryonic Xenopus laevis myocytes.
R Chauhan-Patel and A E Spruce (1998)
J. Physiol. 512, 109-118
   Abstract »    Full Text »    PDF »
A Model for Signal Transduction during Gamete Release in the Fucoid Alga Pelvetia compressa.
G. Anthony Pearson and S. Howard Brawley (1998)
Plant Physiology 118, 305-313
   Abstract »    Full Text »
Modulation of Olfactory Bulb Neuron Potassium Current by Tyrosine Phosphorylation.
D. A. Fadool and I. B. Levitan (1998)
J. Neurosci. 18, 6126-6137
   Abstract »    Full Text »    PDF »
Lasp-1 is a regulated phosphoprotein within the cAMP signaling pathway in the gastric parietal cell.
C. S. Chew, J. A. Parente Jr., C.-J. Zhou, E. Baranco, and X. Chen (1998)
Am J Physiol Cell Physiol 275, C56-C67
   Abstract »    Full Text »    PDF »
Activation of protein tyrosine kinase PYK2 by the m1 muscarinic acetylcholine receptor.
J. S. Felsch, T. G. Cachero, and E. G. Peralta (1998)
PNAS 95, 5051-5056
   Abstract »    Full Text »    PDF »
Evidence for Direct Physical Association between a K+ Channel (Kir6.2) and an ATP-Binding Cassette Protein (SUR1) Which Affects Cellular Distribution and Kinetic Behavior of an ATP-Sensitive K+ Channel.
E. Lorenz, A. E. Alekseev, G. B. Krapivinsky, A. J. Carrasco, D. E. Clapham, and A. Terzic (1998)
Mol. Cell. Biol. 18, 1652-1659
   Abstract »    Full Text »
Modulation of Voltage-dependent Ca2+ Channels in Rabbit Colonic Smooth Muscle Cells by c-Src and Focal Adhesion Kinase.
X.-Q. Hu, N. Singh, D. Mukhopadhyay, and H. I. Akbarali (1998)
J. Biol. Chem. 273, 5337-5342
   Abstract »    Full Text »    PDF »
Growth Factor Receptor Tyrosine Kinases Acutely Regulate Neuronal Sodium Channels through the Src Signaling Pathway.
M. D. Hilborn, R. R. Vaillancourt, and S. G. Rane (1998)
J. Neurosci. 18, 590-600
   Abstract »    Full Text »    PDF »
Expression of Voltage-Gated Potassium Channels Decreases Cellular Protein Tyrosine Phosphorylation.
T. C. Holmes, K. Berman, J. E. Swartz, D. Dagan, and I. B. Levitan (1997)
J. Neurosci. 17, 8964-8974
   Abstract »    Full Text »    PDF »
Modulation of Rod Photoreceptor Cyclic Nucleotide-Gated Channels by Tyrosine Phosphorylation.
E. Molokanova, B. Trivedi, A. Savchenko, and R. H. Kramer (1997)
J. Neurosci. 17, 9068-9076
   Abstract »    Full Text »    PDF »
Modulation of the Kv1.3 Potassium Channel by Receptor Tyrosine Kinases.
M. R. Bowlby, D. A. Fadool, T. C. Holmes, and I. B. Levitan (1997)
J. Gen. Physiol. 110, 601-610
   Abstract »    Full Text »    PDF »
Phosphorylation of the Kv2.1 K+ Channel Alters Voltage-Dependent Activation.
H. Murakoshi, G. Shi, R. H. Scannevin, and J. S. Trimmer (1997)
Mol. Pharmacol. 52, 821-828
   Abstract »    Full Text »
Rescue of osteoclast function by transgenic expression of kinase-deficient Src in src-/- mutant mice.
P. L. Schwartzberg, L. Xing, O. Hoffmann, C. A. Lowell, L. Garrett, B. F. Boyce, and H. E. Varmus (1997)
Genes & Dev. 11, 2835-2844
   Abstract »    Full Text »    PDF »
Cystic fibrosis transmembrane conductance regulator is an epithelial cell receptor for clearance of Pseudomonas aeruginosa from the lung.
G. B. Pier, M. Grout, and T. S. Zaidi (1997)
PNAS 94, 12088-12093
   Abstract »    Full Text »    PDF »
Human Homologue of the Drosophila Discs Large Tumor Suppressor Binds to p56lck Tyrosine Kinase and Shaker Type Kv1.3 Potassium Channel in T Lymphocytes.
T. Hanada, L. Lin, K. G. Chandy, S. S. Oh, and A. H. Chishti (1997)
J. Biol. Chem. 272, 26899-26904
   Abstract »    Full Text »    PDF »
Lysophosphatidylcholine Modulates Cardiac INa via Multiple Protein Kinase Pathways.
C. L. Watson and M. R. Gold (1997)
Circ. Res. 81, 387-395
   Abstract »    Full Text »
Tyrosine Phosphorylation of Connexin 43 by v-Src Is Mediated by SH2 and SH3 Domain Interactions.
M. Y. Kanemitsu, L. W. M. Loo, S. Simon, A. F. Lau, and W. Eckhart (1997)
J. Biol. Chem. 272, 22824-22831
   Abstract »    Full Text »    PDF »
Requirement for Tyrosine Phosphatase during Serotonergic Neuromodulation by Protein Kinase C.
S. Catarsi and P. Drapeau (1997)
J. Neurosci. 17, 5792-5797
   Abstract »    Full Text »    PDF »
Tyrosine Phosphorylation of Nicotinic Acetylcholine Receptor Mediates Grb2 Binding.
M. Colledge and S. C. Froehner (1997)
J. Neurosci. 17, 5038-5045
   Abstract »    Full Text »    PDF »
Simultaneous Binding of Two Protein Kinases to a Calcium-Dependent Potassium Channel.
J. Wang, Y. Zhou, H. Wen, and I. B. Levitan (1999)
J. Neurosci. 19, RC4
   Abstract »    Full Text »    PDF »
Enhanced Activity of a Large Conductance, Calcium-sensitive K+ Channel in the Presence of Src Tyrosine Kinase.
S. Ling, G. Woronuk, L. Sy, S. Lev, and A. P. Braun (2000)
J. Biol. Chem. 275, 30683-30689
   Abstract »    Full Text »    PDF »
A Catalytically Inactive Mutant of Type I cGMP-dependent Protein Kinase Prevents Enhancement of Large Conductance, Calcium-sensitive K+ Channels by Sodium Nitroprusside and cGMP.
R. D. Swayze and A. P. Braun (2001)
J. Biol. Chem. 276, 19729-19737
   Abstract »    Full Text »    PDF »
Isoform-specific Localization of Voltage-gated K+ Channels to Distinct Lipid Raft Populations. TARGETING OF Kv1.5 TO CAVEOLAE.
J. R. Martens, N. Sakamoto, S. A. Sullivan, T. D. Grobaski, and M. M. Tamkun (2001)
J. Biol. Chem. 276, 8409-8414
   Abstract »    Full Text »    PDF »
Regulation of the L-type Calcium Channel by alpha 5beta 1 Integrin Requires Signaling between Focal Adhesion Proteins.
X. Wu, G. E. Davis, G. A. Meininger, E. Wilson, and M. J. Davis (2001)
J. Biol. Chem. 276, 30285-30292
   Abstract »    Full Text »    PDF »
A mechanism for combinatorial regulation of electrical activity: Potassium channel subunits capable of functioning as Src homology 3-dependent adaptors.
M. N. Nitabach, D. A. Llamas, R. C. Araneda, J. L. Intile, I. J. Thompson, Y. I. Zhou, and T. C. Holmes (2001)
PNAS 98, 705-710
   Abstract »    Full Text »    PDF »
Direct inhibition of the cloned Kv1.5 channel by AG-1478, a tyrosine kinase inhibitor.
B. H. Choi, J.-S. Choi, D.-J. Rhie, S. H. Yoon, D. S. Min, Y.-H. Jo, M.-S. Kim, and S. J. Hahn (2002)
Am J Physiol Cell Physiol 282, C1461-C1468
   Abstract »    Full Text »    PDF »


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