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BK Ca Channels Activating at Resting Potential without Calcium in LNCaP Prostate Cancer Cells

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Abstract

Large-conductance Ca2+-dependent K+ (BKCa) channels are activated by intracellular Ca2+ and membrane depolarization in an allosteric manner. We investigated the pharmacological and biophysical characteristics of a BKCa-type K+ channel in androgen-dependent LNCaP (lymph node carcinoma of the prostate) cells with novel functional properties, here termed BKL. K+ selectivity, high conductance, activation by Mg2+ or NS1619, and inhibition by paxilline and penitrem A largely resembled the properties of recombinant BKCa channels. However, unlike conventional BKCa channels, BKL channels activated in the absence of free cytosolic Ca2+ at physiological membrane potentials; the half-maximal activation voltage was shifted by about −100 mV compared with BKCa channels. Half-maximal Ca2+-dependent activation was observed at 0.4 μM for BKL (at −20 mV) and at 4.1 μM for BKCa channels (at +50 mV). Heterologous expression of hSlo1 in LNCaP cells increased the BKL conductance. Expression of hSlo-β1 in LNCaP cells shifted voltage-dependent activation to values between that of BKL and BKCa channels and reduced the slope of the P open (open probability)-voltage curve. We propose that LNCaP cells harbor a so far unknown type of BKCa subunit, which is responsible for the BKL phenotype in a dominant manner. BKL-like channels are also expressed in the human breast cancer cell line T47D. In addition, functional expression of BKL in LNCaP cells is regulated by serum-derived factors, however not by androgens.

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References

  • Anwer K., Toro L., Oberti C., Stefani E., Sanborn B M. 1992. Ca2+-activated K+ channels in pregnant rat myometrium: modulation by a β-adrenergic agent. Am. J. Physiot. 263:1049–1056

    Google Scholar 

  • Basrai D., Kraft R., Bollensdorfff C., Liebmann L., Benndorf K., Patt S. 2002. BK channel blockers inhibit potassium-induced proliferation of human astrocytoma cells Neuro Report. 13:403–407

    CAS  Google Scholar 

  • Brenner R., Perez G.J., Bonev A.D., Eckman D.M., Kosek J.C, Wiler S.W., Patterson A.J., Nelson M.T., Aldrich R.W. 2000. Vasoregulation by the β1 subunit of the calcium-activated potassium channel. Nature 407:870–876

    Article  PubMed  CAS  Google Scholar 

  • Diaz L., Meera P., Amigo J., Stefani E., Alvarez O., Toro L., Latorre R. 1998. Role of the S4 segment in a voltage-dependent calcium-sensitive potassium (hSlo) channel. J. Biol. Chem. 273:32430–32436

    Article  PubMed  Google Scholar 

  • Dworetzky S.I., Boissard C.G., Lum-Ragan J.T., McKay M.C., Post-Munson D.J., Trojnacki. J.T., Chang C.-P., Gribkoff V.K. 1996 Phenotypic alteration of a human BK (hSlo) channel by hSloβ subunit coexpression; changes in blocker sensitivity, activation/relaxation and inactivation kinetics, and protein kinase A modulation. J. Neurosci. 16:4543–4550

    PubMed  CAS  Google Scholar 

  • Erxleben C., Everhart A.L., Romeo C., Florence H., Bauer M.B., Alcorta DA., Rossie S., Shipston M.J., Armstrong D.L. 2002. Interacting effects of N-terminal variation and strex exon splicing on slo potassium channel regulation by calcium, phosphorylation. and oxidation. J. Biol. Chem. 277: 27045–27052

    Article  PubMed  CAS  Google Scholar 

  • Fernández-Fernández J.M., Tomás M., Vázquez E., Orio P., Lalorre R., Senti M., Marrugsit J., Valverde M.A. 2004. Gain-of-function mutation in the KCNMB1 potassium channel subunit is associated with low prevalence of diastolic hypertension. Clin. Invest. 113:1032–1039

    Article  PubMed  CAS  Google Scholar 

  • Gribkoff V.K., Starret Jr. J.E., Dworetzky S.I. 1997. The pharmacology and molecular biology of large-conductance calcium-activated (BK) potassium channels. Adv. Pharmacol. 37:319–348

    Article  PubMed  CAS  Google Scholar 

  • Gutiérrez A.A., Arias J.M., Garcia L., Mas-Oliva J., Guerrero-Hernandéz A. 1999. Activation of a Ca2+-permeable cation channel by two different inducers of apoptosis in a human prostatic cancer cell line. J. Physiol. 517:95–107

    Article  PubMed  Google Scholar 

  • Horoszewicz J.S., Leong S.S, Kawinski E., Karr J.P., Rosenthal H., Chu T.M., Mirand E.A., Murphy G.P. 1963. LNCaP model of human prostatic carcinoma. Cancer Res 43:1908–1916

    Google Scholar 

  • Horrigan F.T., Cur J., Aldnch R.W. 1999. Allosteric voltage gating of potassium channels I: mSlo ionic currents in the absence of Ca2+. J. Gen. Physiol. 114:277–304

    Article  PubMed  CAS  Google Scholar 

  • Jin P., Weiger T.M., Wu Y., Levitan I.B. 2002. Phosphorylation-dependent functional coupling of hSlo calcium-dependent potassium channel and its hβ4 subunit. J. Biol Chem. 277:10014–10020

    Article  PubMed  CAS  Google Scholar 

  • Kaczorowski G.J., Knaus H.-G., Leonard R.J., McManus O.B., Garcia M.L. 1996. High-conductance calcium-activated potassium channels: structure, pharmacolcgy, and function. J Bioenerg. Biomembr. 28:255–267

    Article  PubMed  CAS  Google Scholar 

  • Knaus H.-G., McManus O.B., Lee S.H., Schmalhofer W.A., Garcia-Calvo M., Helms L.M.H., Sanchez M., Giangiacomo K., Reuben J.P., Smith A.B., 3rd, Kaczorowski G.J., Garcia M.L. 1994. Tremorgenic indole alkaloids potently inhibit smooth muscle high-conductance calcium-activated potassium channels. Biochemistry 33:5819–5828

    Article  PubMed  CAS  Google Scholar 

  • Kraft R., Benndorf K., Patt S. 2000. Large conductance Ca2+ activated K+ channels in human meningioma cells. J. Membrane. Biol. 175:25–33

    Article  CAS  Google Scholar 

  • Lai G.-J., McCobb D.P. 2002. Opposing actions of adrenal androgens and glucocorticoids on alternative splicing of Slo potassium channels in bovine chromaffin cells. Proc. Natl. Acact. Set. USA 99:7722–7727

    Article  PubMed  CAS  Google Scholar 

  • Lippiat J.D., Standen N.B., Davies. N.W. 2000. A residue in the intracellular vestibule of the pore is critical for gating and permeation in Ca2+-activated K+ (BKCa) channels. J Physiot. 529:131–138

    Article  PubMed  CAS  Google Scholar 

  • Lippiat J.D., Standen N.B., Harrow I.D., Phillips S.C., Davies N.W. 2003. Properties of BKCa channels formed by bicistronic expression of hSlo α and β1-4 subunits in HEK293 cells. J. Membrane. Biol. 192:141–148

    Article  CAS  Google Scholar 

  • Liu X., Chang Y., Reinhart P.M., Sontheimer H. 2002. Cloning and characterization of glioma BK, a novel BK channel isoform highly expressed in human glioma cells. J. Neurosci. 22:1640–1849

    PubMed  CAS  Google Scholar 

  • Marrion M.V., Tavalin S.J. 1998. Selective activation of Ca2+-activated K+ channels by co-localized Ca2+ channels in hippocampal neurons. Nature 395:900–905

    Article  PubMed  CAS  Google Scholar 

  • Meera P., Wallner M., Toro L. 2000. A neuronal β subunit (hKCNMB4) makes the large conductance, voltage- and Ca2+-activated K+ channel resistant to charybdotoxin and iberiotion. Proc. Natl. Acad. Sci. USA. 97:5562–5567

    Article  PubMed  CAS  Google Scholar 

  • Meera P., Wallner M., Jiang Z., Toro L. 1996. A calcium switch for the functional coupling between (hslo) and β subunits (KV,Caβ) of maxi K channels. FEBS Lett. 382:84–38

    Article  PubMed  CAS  Google Scholar 

  • Meera P., Wallner M., Song M., Toro L. 1997. Large conductance voltage- and calcium-dependent K+ channel, a distinct member of voltage-dependention channels with seven N-terrninal transmembrane segments (S0-S6), an extracellular N terminus, and an intracellular (S9-S10) C terminus. Proc. Natl Acad. Set. USA 94:14066–14071

    Article  PubMed  CAS  Google Scholar 

  • Orio P., Rojas P., Ferreira G., Latorre R. 2002. New disguises for an old channel. MaxiK channel β-subunits. News Physiol. Sci. 17:156–161

    PubMed  CAS  Google Scholar 

  • Ramanathan K., Michael T.H., Jiang G.-J., Hiel H., Fuchs P.A. 1999. A molecular mechanism for electrical tuning of cochleae hair cells. Science 283:215–217

    Article  PubMed  CAS  Google Scholar 

  • Sansom S.C., Stockand J.D. 1994. Differential Ca2+ sensitivities of BK(Ca) isochannels in bovine mesenteric vascular smooth muscle. Am. J. Physiol. 266:1182–1189

    Google Scholar 

  • Santarelli L.C., Chen J., Hsinernann S.H., Hoshi T. 2004. The β1 subunit enhances oxidative regulation of large-conductance calcium -activated K+ channels J. Gen. Physiol. 124:357–370

    Article  PubMed  CAS  Google Scholar 

  • Santarelli, L.C., Wassef, R., Heinemann, S.H., Hoshi, I. 2006. Three methionine residues located within the regulator of conductance for K+ (RCK) domains confer oxidative sensitivity to large-conductance Ca2+- activated K+ channels. J. Physiol. 571:329–359

    Article  PubMed  CAS  Google Scholar 

  • Schubert R., Nelson M.T. 2001. Protein kinases: tuners of the BKCa channel in smooth musde. Trends Pharmacol. 22:505–512

    Article  PubMed  CAS  Google Scholar 

  • Shao L.-R., Halvorsrud R., Borg-Graham L., Storm J.F. 1999. The role of BK-type Ca2+-dependent K+ channels in spike broadening during repetitive firing in rat hippocampal pyramidal cells. J. Physiol. 521:135–146

    Article  PubMed  CAS  Google Scholar 

  • Shen K.-Z., Lagrutta A., Davies N., Standen N., Adeiman J., North R. 1994. Tetraethylammonium block of Slowpoke calcium -activated potassium channels expressed in Xenopus oocytes: evidence for tetrameric channel formation. Pfluegures Arch. 426:440–445

    Article  CAS  Google Scholar 

  • Shi J., Cui J. 2001. Intracellular Mg2+ enhances the function of BK-type Ca2+-activated K+ channels. J. Gen. Physiol. 113:589–605

    Article  PubMed  CAS  Google Scholar 

  • Skryrna R., van Coppenolle P., Dufy-Barbe L., Dufy B., Prevarskaya N. 1999. Characterization of Ca2+-inhibited potassium channels in the LNCaP human prostate cancer cell line. Receptors Channels 6:241–253

    PubMed  CAS  Google Scholar 

  • Strøbaeck D., Christopherson P., Holm N.R., Moldt P., Ahring P.K. Johgnsen T.E., Olesen S.-P. 1996. Modulation of the Ca2+-dependenl K+ channel hslo, by the substituted diphenylurea MS 1608, paxilline and internal Ca2+. Neuropharmacology 35:903–914

    Article  PubMed  Google Scholar 

  • Thalmann. G.N., Anezinis P.E., Chang S.M., Zhau H.E., Kirn E.E., Hopwood V.L., Pathak S., von Eschenbach A.C., Chung L.W. 1994. Androgen-independent cancer progression and bone metastasis in the LNCaP model of human prostate cancer. Cancer Res. 54:2577–2531

    PubMed  CAS  Google Scholar 

  • Thurm H., Fakler B., Oliver D. 2005 Ca2+-independent activation of BKCa channels at negative potentials in mammalian inner hair cells J. Physiol 569:137–151

    Article  CAS  Google Scholar 

  • Tseng-Crank J., Foster C.D., Krause J.D., Mertz R., Godinot N., DiChiara T.J., Reinhart P.H. 1994. Cloning, expression, and distribution of functionally distinct Ca2+-activated K+ channel isoforms from: human brain. Neuron 13:1315–1330

    Article  PubMed  CAS  Google Scholar 

  • Wang Y.-W., Ding J.P., Xia X.-M., Lingie C.J. 2002. Consecquences of the stoichiometry of Slo1 α and auxiliary β subunits on functional properties of large-conductance Ca2+-activated K+ channels. J. Neurosci. 22:1550–1561

    PubMed  CAS  Google Scholar 

  • Weiger T.M., Hermann A., Levitan I.B. 2002. Modulation of calcium-activated potassium channels. J. Comp. Physiol A. 188:79–87

    Article  CAS  Google Scholar 

  • Weaver A.K., Llu X., Sontheimer H. 2004. Role for calcium-activated potassium channels (BK) in growth control of human malignant glioma J. Neurrosci. Res. 78:224–234

    Article  PubMed  CAS  Google Scholar 

  • Xia X.-M., Zeng X., Lingle. C.J. 2002. Multiple regulatory sites in large-conductance calcium-activated potassium channels. Nature 418:880–884

    Article  PubMed  CAS  Google Scholar 

  • Xie J., McCobb D.P. 1998. Control of alternative splicing of potassium channels toy hormones. Science 280:443–446

    Article  PubMed  CAS  Google Scholar 

  • Zhau H.V., Chang S.M., Chen B.Q., Wang Y., Zhang H., Kao C., Sang Q.A., Pathak S.J., Chung L.W. 1996. Androgen-reprassed phenotype in human prostate cancer. Proc. Natl. Acad. Sci. USA 93:15152–15157

    Article  PubMed  CAS  Google Scholar 

  • Zhou X.-B., Arntz C., Kamm S., Motejlek K., U., Wang G.-X., Ruth P., Korth M. 2001. A molecular switch for specific simulation of the BKCa channel by cGMP and cAMP kinase. J. Biol. Chem. 276:43239–43245

    Article  PubMed  CAS  Google Scholar 

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Acknowledgement

We are grateful for technical assistance by S. Arend and A. Rossner and for helpful discussions with R. Schönherr. This work was supported by the DFG (HE 2993/2), TMWFK (B378-01027) and National Institute of Health.

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Authors and Affiliations

  1. Institute of Molecular Cell Biology, Molecular and Cellular Biophysics, Friedrich Schiller University , Jena, Germany

    G. Gessner, K. Schönherr, M. Soom, A. Hansel & S.H. Heinemann

  2. Institute of Human Genetics and Anthropology, Molecular Genetics, Friedrich Schiller University , Jena, Germany

    M. Asim & A. Baniahmad

  3. Center for Anatomy, Electronmicroscopy and Molecular Neuroanatomy, Charité Berlin, Germany

    C. Derst

  4. Department of Physiology, University of Pennsylvania, Philadelphia, PA, USA

    T. Hoshi

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  1. G. Gessner
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  2. K. Schönherr
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  9. S.H. Heinemann
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Correspondence to S.H. Heinemann.

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Gessner, G., Schönherr, K., Soom, M. et al. BK Ca Channels Activating at Resting Potential without Calcium in LNCaP Prostate Cancer Cells. J Membrane Biol 208, 229–240 (2006). https://doi.org/10.1007/s00232-005-0830-z

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  • DOI: https://doi.org/10.1007/s00232-005-0830-z

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