Skip to main content
Log in

Calcium-independent K+-selective channel from chromaffin granule membranes

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Intact adrenal chromaffin granules and purified granule membrane ghosts were allowed to fuse with acidic phospholipid planar bilayer membranes in the presence of Ca2+ (1 mm). From both preparations, we were able to detect a large conductance potassium channel (ca. 160 pS in symmetrical 400 mm K+), which was highly selective for K+ over Na+ (P k/P Na = 11) as estimated from the reversal potential of the channel current. Channel activity was unaffected by charybdotoxin, a blocker of the [Ca2+] activated K+ channel of large conductance. Furthermore, this channel proved quite different from the previously described channels from other types of secretory vesicle preparations, not only in its selectivity and conductance, but also in its insensitivity to both calcium and potential across the bilayer. We conclude that the chromaffin granule membrane contains a K+-selective channel with large conductance. We suggest that the role of this channel may include ion movement during granule assembly or recycling, and do not rule out events leading to exocytosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Anderson, C.S., MacKinnon, R., Smith, C., Miller, C. 1988. Charybdotoxin block of single Ca2+-activated K+-channels. Effects of channel gating, voltage, and ionic strength. J. Gen. Physiol. 91:317–333

    Google Scholar 

  • Anton, A.H., Sayre, D.F. 1962. A study of the factor affecting the aluminum oxide-trihydroxyindole procedure for the analysis of catecholamines. J. Pharm. Exptl. Ther. 138:360–375

    Google Scholar 

  • Baker, P.F., Knight, D.E. 1981. Calcium control of exocytosis and endocytosis in bovine adrenal medullary cells. Philos. Trans. R. Soc. B. 296:83–103

    Google Scholar 

  • Bergmeyer, H.U. 1974. Methods of Enzymatic Analysis. 1:495–496. Academic, New York

    Google Scholar 

  • Blair, H.C., Schlesinger, P.H. 1990. Purification of a stilbene sensitive chloride channel and reconstitution of chloride conductivity into phospholipid vesicles. Biochem. Biophys. Res. Commun. 171:920–925

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Breckenridge, L.J., Almers, W. 1987. Final steps in exocytosis observed in a cell with giant secretory granules. Proc. Natl. Acad. Sci. USA 84:1945–1949

    Google Scholar 

  • Brocklehurst, K.W., Pollard, H.B. 1988a. Osmotic effects in membrane fusion during exocytosis. Curr. Topics in Membr. Transp. 32:203–225

    Google Scholar 

  • Brocklehurst, K.W., Pollard, H.B. 1988b. Chemiosmotic events. In: Energetics of Secretion Responses, J.-W.N. Akkerman, editor, 2:47–61. CRC Press, Boca Raton

    Google Scholar 

  • Brocklehurst, K.W., Pollard, H.B. 1990. Cell Biology of Secretion. In: Peptide Hormone Secretion. A Practical Approach. J.C. Hutton and K. Siddle, editors. pp. 233–255. IRL/Oxford, Oxford, New York, Tokyo

    Google Scholar 

  • Creutz, C.E., Pollard, H.B. 1980. A biophysical model of the chromaffin granule: accurate description of the kinetics of ATP and Cl-dependent granule lysis. Biophys. J. 31:255–270

    Google Scholar 

  • De Riemer, S.A., Rahamimoff, R., Sakmann, B., Stadler, H. 1987. Conductances and channels in fused synaptic vesicles from Torpedo electric organ. J. Physiol. 368:84P

    Google Scholar 

  • Fenwick, E.M., Marty, A., Neher, E. 1982. A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcho line. J. Physiol. 331:557–597

    Google Scholar 

  • Finkelstein, A., Zimmerberg, J., Cohen, F.S. 1986. Osmotic swelling of vesicles: its role in the fusion of vesicles with planar phospholipid bilayer membranes and its possible role in exocytosis. Annu. Rev. Physiol. 48:163–174

    Google Scholar 

  • Holz, R.W. 1979. Measurement of membrane potential of chromaffin granules by accumulation of triphenylmethylphosphonium cation. J. Biol. Chem. 254:6703–6709

    Google Scholar 

  • Kelner, K.L., Levine, R.A., Monta, K., Pollard, H.B. 1985. A comparison of tryhydroxyindole and HPLC electrochemical methods for catecholamine measurement in adrenal chromaffin cells. Neurochem. Int. 7:373–378

    Google Scholar 

  • Knaus, P., Marqueze-Pouey, B., Scherer, H., Betz, H. 1990. Synaptoporin, a novel putative channel protein of synaptic vesicles. Neuron 5:453–462

    Google Scholar 

  • Lee, C.J., Dayanithi, G., Nordmann, J.J., Lemos, J.R. 1992. Possible role during exocytosis of a Ca2+-activated channel in neurohypophysial granules. Neuron 8:335–342

    Google Scholar 

  • Lemos, J.R., Ocorr, K.A., Nordmann, J.J. 1989a. Possible role for ionic channels in neurosecretory granules of the rat neurohypophysis. In: Secretion and its Control. C. Armstrong and G. Oxford, editors. pp. 334–347. Rockefeller University, New York

    Google Scholar 

  • Lemos, J.R., Ocorr, K.A., Nordmann, J.J. 1989b. Possible role for ionic channels in neurosecretory granules of the rat neurohypophysis. Soc. Gen. Physiol. Ser. 44:333–347

    Google Scholar 

  • Lindau, M. 1991. Time-resolved capacitance measurements: monitoring exocytosis in single cells. Quart. Rev. Biophys. 24:75–101

    Google Scholar 

  • Lingg, G., Fischer-Colbrie, R., Schmidt, W., Winkler, H. 1983. Exposure of an antigen of chromaffin granules on cell surface during exocytosis. Nature 301:610–611

    Google Scholar 

  • McCaman, R.E., McCamen, M.W., Hunt, J.M., Smith, M.S. 1965. Microdetermination of monoamine oxidase and 5-hydroxytryptophan decarboxylase activities in nervous tissues. J. Neurochem. 12:15–23

    Google Scholar 

  • MacKinnon, R., Miller, C. 1988. Mechanism of Charybdotoxin block of the high-conductance Ca2+-activated K+-channel. J. Gen. Physiol. 91:335–349

    Google Scholar 

  • Nagatsu, T., Udenfriend, S. 1972. Photometric assay of dopamine-b-hydroxylase activity in human blood. Clin. Chem. 18:980–983

    Google Scholar 

  • Neher, E., Marty, A. 1982. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc. Natl. Acad. Sci. USA 79:6712–6716

    Google Scholar 

  • Obendorf, D., Schwarzenbrunner, U., Fischer-Colbrie, R., Laslop, A., Winkler, H. 1988. In adrenal medulla synaptophysin (protein p38) is present in chromaffin granules and in a special vesicle population. J. Neurochem. 51:1573–1580

    Google Scholar 

  • Ornberg, R.L., Kuijpers, G.A., Leapman, R.D. 1988. Electron probe microanalysis of the subcellular compartments of bovine adrenal chromaffin cells. J. Biol. Chem. 263:1488–1493

    Google Scholar 

  • Ornberg, R.L., Reese, T.S. 1981. Beginning of exocytosis captured by rapid-freezing of limulus amoebocytes. J. Cell Biol. 90:40–54, International Rev. Cytol. 58:159–197

    Article  Google Scholar 

  • Pazoles, C.J., Claggett, C.E., Creutz, C.E., Pollard, H.B., Weinbach, E.C. 1980. Identification and subcellular localization of catalase activity in bovine adrenal medulla and cortex. Arch. Bioch. Biophys. 200:434–443

    Google Scholar 

  • Pazoles, C.J., Pollard, H.B. 1978. Evidence for activation of anion transport in ATP-evoked transmitter release from isolated secretory vesicles. J. Biol. Chem. 253:3962–3969

    Google Scholar 

  • Penner, R., Neher, E. 1989. The patch-clamp technique in the study of secretion. Trends Neurosci. 12:159–163

    Google Scholar 

  • Picaud, S., Marty, A., Trautmann, A., Grynszpan-Winograd, O., Henry, J.-P. 1984. Incorporation of chromaffin granule membranes into large-size vesicles suitable for patch-clamp recording. FEBS Lett. 178:20–24

    Google Scholar 

  • Pollard, H.B., Zinder, O., Hoffman, P.G., Nikodijevik, O. 1976. Regulation of the transmembrane potential of isolated chromaffin granules by ATP, ATP analogs, and external pH. J. Biol. Chem. 251:4544–4545

    Google Scholar 

  • Pollard, H.B., Menard, R., Brandt, H.A., Pazoles, C.J., Creutz, C.E., Ramu, A. 1978. Application of Bradford's protein assay to adrenal gland subcellular fractions. Anal. Bioch. 86:761–763

    Google Scholar 

  • Pollard, H.B., Pazoles, C.J., Creutz, C.E., Zinder, O. 1979a. The chromaffin granule and possible mechanisms of exocytosis. International Rev. Cytol. 58:159–197

    Google Scholar 

  • Pollard, H.B., Shindo, H., Creutz, C.E., Zinder, O. 1979b. Internal pH and state of ATP in adrenergic chromaffin granules determined by 31P nuclear magnetic resonance spectroscopy. J. Biol. Chem. 254:1170–1177

    Google Scholar 

  • Pollard, H.B., Pazoles, C.J., Creutz, C.E. 1980. Evidence in support of a chemiosmotic mechanism for exocytosis from platelets, parathyroid and chromaffin cells. In: Catecholamines: Basic and Clinical Frontiers. Editors I. Hopin and E. Usdin, editors. 1:328–330. Pergamon, Oxford

    Google Scholar 

  • Pollard, H.B., Pazoles, C.J., Creutz, C.E., Scott, J.H., Zinder, O., Hotchkiss, A. 1984. An osmotic mechanism for exocytosis from dissociated chromaffin cells. J. Biol. Chem. 259:1114–1121

    Google Scholar 

  • Pollard, H.B., Rojas, E. 1988. Calcium-activated synexin forms highly selective, voltage-gated channels in phosphatidylserine bilayer membranes. Proc. Natl. Acad. Sci. USA 85:2974–2978

    Google Scholar 

  • Pollard, H.B., Bums, A.L., Rojas, E. 1990. Synexin (annexin VII): A cytosolic calcium-binding protein which promotes membrane fusion and forms calcium channels in artificial bilayers and natural membranes. J. Membrane Biol. 117:101–112

    Google Scholar 

  • Rahamimoff, R., De Riemer, S.A., Sakmann, B., Stadler, H., Yakir, N. 1988. Ion channels in synaptic vesicles from Torpedo electric organ. Proc. Natl. Acad. Sci. USA 85:5310–5314

    Google Scholar 

  • Rahamimoff, R., De Riemer, S.A., Ginsburg, S., Kaiserman, I., Sakmann, B., Stadler, H., Yakir, N. 1989. Ionic channels in synaptic vesicles: are they involved in transmitter release? Quarterly J. Exp. Physiol. 74:1019–1031

    Google Scholar 

  • Rahamimoff, R., De Riemer, S.A., Ginsburg, S., Kaiserman, I., Sakmann, B., Stadler, H., Yakir, N. 1990. Ionic channels and proteins in synaptic vesicles: facts and speculations. J. Basic Clin. Physiol. Pharmacol. 1:7–17

    Google Scholar 

  • Sakmann, B., Neher, E. 1983. Single-channel Recording. Plenum, New York

    Google Scholar 

  • Salama, G., Johnson, R.G., Scarpa, A. 1980. Spectrophotometric measurements of transmembrane potential and pH gradients in chromaffin granules. J. Gen. Physiol. 75:109–140

    Google Scholar 

  • Stanley, E.F., Ehrenstein, G. 1985. A model for exocytosis based on the opening of calcium-activated potassium channels in vesicles. Life Sci. 25:1985–1995

    Google Scholar 

  • Stanley, E.F., Ehrenstein, G., Russell, J.T. 1988. Evidence for anion channels in secretory vesicles. Neuroscience 25: 1035–1039

    Google Scholar 

  • Trifaro, J.M., Poisner, A.M. 1982. Common properties in the mechanisms of synthesis, processing and storage of secretory products. In: The Secretory Granule. A.M. Poisner and J.M. Trifaro, editors. Elsevier Biomedical

  • Thomas, L., Hartung, K., Langosch, D., Rehm, H., Bamberg, E., Franke, W., Betz, H. 1988. Identification of synaptophysin as a hexameric channel protein of the synaptic vesicle membrane. Science 242:1050–1053

    Google Scholar 

  • Zinder, O., Menard, R., Lovenberg, W., Pollard, H.B. 1977. Direct evidence for co-localization of adenylate cyclase, dopamine-beta-hydroxylase and cytochrome b562 to bovine chromaffin granule membranes. Biochem. Biophys. Res. Comm. 79:707–712

    Google Scholar 

  • Zinder, O., Nikodijevic, O., Hoffman, P.G., Pollard, H.B. 1976. Regulation of secretion from the adrenal medulla. Evidence for adenylate cyclase activity in secretory vesicle membranes. J. Biol. Chem. 251:2179–2181

    Google Scholar 

  • Zinder, O., Hoffman, P.G., Bonner, W.M., Pollard, H.B. 1978. Comparison of chemical properties of purified plasma membranes and secretory vesicle membranes from the bovine adrenal medulla. Cell. Tiss. Res. 188:153–170

    Google Scholar 

  • Von Grafenstein, H., Roberts, C.S., Baker, P.F. 1986. The kinetics of the exocytosis endocytosis secretory cyclein bovine adrenal medullary cells. J. Cell Biol. 103:2343–2352

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arispe, N., Pollard, H.B. & Rojas, E. Calcium-independent K+-selective channel from chromaffin granule membranes. J. Membarin Biol. 130, 191–202 (1992). https://doi.org/10.1007/BF00231896

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00231896

Key Words

Navigation