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. 1988 May;7(5):1449–1454. doi: 10.1002/j.1460-2075.1988.tb02962.x

Incorporation in lipid bilayers of a large conductance cationic channel from mitochondrial membranes.

M Thieffry 1, J F Chich 1, D Goldschmidt 1, J P Henry 1
PMCID: PMC458395  PMID: 2457497

Abstract

Membranes from subcellular fractions of adrenal medulla were incorporated in phospholipid bilayers formed at the tip of microelectrodes. Current fluctuations recorded in the presence of a transmembrane potential revealed the existence of a voltage-dependent channel of large conductance. This channel is characterized by fast kinetics and four conductance levels separated by jumps of 100, 220 and 220 pS in 150 mM NaCl. It is permeant to Na+,K+, tetraethylammonium, Cl- and acetate and has some cation selectivity. Exposure to trypsin or pronase abolished the voltage-dependence. Upon subcellular fractionation, the activity was found to be associated with mitochondria. A similar activity was observed in mitochondrial fractions from other organs. By its kinetics, its selectivity and its potential-dependence, this channel differs from the voltage-dependent anion channel of outer mitochondrial membranes.

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  1. APPELMANS F., WATTIAUX R., DE DUVE C. Tissue fractionation studies. 5. The association of acid phosphatase with a special class of cytoplasmic granules in rat liver. Biochem J. 1955 Mar;59(3):438–445. doi: 10.1042/bj0590438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anner B., Mossmayer M. Rapid determination of inorganic phosphate in biological systems by a highly sensitive photometric method. Anal Biochem. 1975 May 12;65(1-2):305–309. doi: 10.1016/0003-2697(75)90514-x. [DOI] [PubMed] [Google Scholar]
  3. Barrett J. N., Magleby K. L., Pallotta B. S. Properties of single calcium-activated potassium channels in cultured rat muscle. J Physiol. 1982 Oct;331:211–230. doi: 10.1113/jphysiol.1982.sp014370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bezanilla F. A high capacity data recording device based on a digital audio processor and a video cassette recorder. Biophys J. 1985 Mar;47(3):437–441. doi: 10.1016/S0006-3495(85)83935-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blatz A. L., Magleby K. L. Single voltage-dependent chloride-selective channels of large conductance in cultured rat muscle. Biophys J. 1983 Aug;43(2):237–241. doi: 10.1016/S0006-3495(83)84344-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  7. Brdiczka D., Knoll G., Riesinger I., Weiler U., Klug G., Benz R., Krause J. Microcompartmentation at the mitochondrial surface: its function in metabolic regulation. Adv Exp Med Biol. 1986;194:55–69. doi: 10.1007/978-1-4684-5107-8_5. [DOI] [PubMed] [Google Scholar]
  8. Carty S. E., Johnson R. G., Scarpa A. The isolation of intact adrenal chromaffin granules using isotonic Percoll density gradients. Anal Biochem. 1980 Aug;106(2):438–445. doi: 10.1016/0003-2697(80)90545-x. [DOI] [PubMed] [Google Scholar]
  9. Colombini M. Purification of VDAC (voltage-dependent anion-selective channel) from rat liver mitochondria. J Membr Biol. 1983;74(2):115–121. doi: 10.1007/BF01870500. [DOI] [PubMed] [Google Scholar]
  10. Coronado R., Latorre R. Phospholipid bilayers made from monolayers on patch-clamp pipettes. Biophys J. 1983 Aug;43(2):231–236. doi: 10.1016/S0006-3495(83)84343-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Doring C., Colombini M. Voltage dependence and ion selectivity of the mitochondrial channel, VDAC, are modified by succinic anhydride. J Membr Biol. 1985;83(1-2):81–86. doi: 10.1007/BF01868740. [DOI] [PubMed] [Google Scholar]
  13. ELLMAN G. L., COURTNEY K. D., ANDRES V., Jr, FEATHER-STONE R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961 Jul;7:88–95. doi: 10.1016/0006-2952(61)90145-9. [DOI] [PubMed] [Google Scholar]
  14. EMMELOT P., BOS C. J., BENEDETTI E. L., RUEMKE P. STUDIES ON PLASMA MEMBRANES. I. CHEMICAL COMPOSITION AND ENZYME CONTENT OF PLASMA MEMBRANES ISOLATED FROM RAT LIVER. Biochim Biophys Acta. 1964 Jul 15;90:126–145. doi: 10.1016/0304-4165(64)90125-4. [DOI] [PubMed] [Google Scholar]
  15. Ewald D. A., Williams A., Levitan I. B. Modulation of single Ca2+-dependent K+-channel activity by protein phosphorylation. Nature. 1985 Jun 6;315(6019):503–506. doi: 10.1038/315503a0. [DOI] [PubMed] [Google Scholar]
  16. Freitag H., Neupert W., Benz R. Purification and characterisation of a pore protein of the outer mitochondrial membrane from Neurospora crassa. Eur J Biochem. 1982 Apr;123(3):629–636. doi: 10.1111/j.1432-1033.1982.tb06578.x. [DOI] [PubMed] [Google Scholar]
  17. Garlid K. D., Beavis A. D. Evidence for the existence of an inner membrane anion channel in mitochondria. Biochim Biophys Acta. 1986;853(3-4):187–204. doi: 10.1016/0304-4173(87)90001-2. [DOI] [PubMed] [Google Scholar]
  18. Labarca P. P., Miller C. A K+-selective, three-state channel from fragmented sarcoplasmic reticulum of frog leg muscle. J Membr Biol. 1981;61(1):31–38. doi: 10.1007/BF01870750. [DOI] [PubMed] [Google Scholar]
  19. Labarca P., Wolff D., Soto U., Necochea C., Leighton F. Large cation-selective pores from rat liver peroxisomal membranes incorporated to planar lipid bilayers. J Membr Biol. 1986;94(3):285–291. doi: 10.1007/BF01869724. [DOI] [PubMed] [Google Scholar]
  20. Latorre R., Miller C. Conduction and selectivity in potassium channels. J Membr Biol. 1983;71(1-2):11–30. doi: 10.1007/BF01870671. [DOI] [PubMed] [Google Scholar]
  21. Mannella C. A. Mitochondrial outer membrane channel (VDAC, porin) two-dimensional crystals from Neurospora. Methods Enzymol. 1986;125:595–610. doi: 10.1016/s0076-6879(86)25048-x. [DOI] [PubMed] [Google Scholar]
  22. Meyer D. I., Burger M. M. Isolation of a protein from the plasma membrane of adrenal medulla which binds to secretory vesicles. J Biol Chem. 1979 Oct 10;254(19):9854–9859. [PubMed] [Google Scholar]
  23. Mihara K., Blobel G., Sato R. In vitro synthesis and integration into mitochondria of porin, a major protein of the outer mitochondrial membrane of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7102–7106. doi: 10.1073/pnas.79.23.7102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miller C. Integral membrane channels: studies in model membranes. Physiol Rev. 1983 Oct;63(4):1209–1242. doi: 10.1152/physrev.1983.63.4.1209. [DOI] [PubMed] [Google Scholar]
  25. Miller C. Voltage-gated cation conductance channel from fragmented sarcoplasmic reticulum: steady-state electrical properties. J Membr Biol. 1978 Apr 20;40(1):1–23. doi: 10.1007/BF01909736. [DOI] [PubMed] [Google Scholar]
  26. Peters T. J., Müller M., De Duve C. Lysosomes of the arterial wall. I. Isolation and subcellular fractionation of cells from normal rabbit aorta. J Exp Med. 1972 Nov 1;136(5):1117–1139. doi: 10.1084/jem.136.5.1117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schein S. J., Colombini M., Finkelstein A. Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from paramecium mitochondria. J Membr Biol. 1976 Dec 28;30(2):99–120. doi: 10.1007/BF01869662. [DOI] [PubMed] [Google Scholar]
  28. Scherman D., Jaudon P., Henry J. P. Characterization of the monoamine carrier of chromaffin granule membrane by binding of [2-3H]dihydrotetrabenazine. Proc Natl Acad Sci U S A. 1983 Jan;80(2):584–588. doi: 10.1073/pnas.80.2.584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Smith A. D., Winkler H. A simple method for the isolation of adrenal chromaffin granules on a large scale. Biochem J. 1967 May;103(2):480–482. doi: 10.1042/bj1030480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith J. S., Coronado R., Meissner G. Sarcoplasmic reticulum contains adenine nucleotide-activated calcium channels. Nature. 1985 Aug 1;316(6027):446–449. doi: 10.1038/316446a0. [DOI] [PubMed] [Google Scholar]
  31. Spray D. C., Saez J. C., Brosius D., Bennett M. V., Hertzberg E. L. Isolated liver gap junctions: gating of transjunctional currents is similar to that in intact pairs of rat hepatocytes. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5494–5497. doi: 10.1073/pnas.83.15.5494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Suarez-Isla B. A., Orozco C., Heller P. F., Froehlich J. P. Single calcium channels in native sarcoplasmic reticulum membranes from skeletal muscle. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7741–7745. doi: 10.1073/pnas.83.20.7741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Suarez-Isla B. A., Wan K., Lindstrom J., Montal M. Single-channel recordings from purified acetylcholine receptors reconstituted in bilayers formed at the tip of patch pipets. Biochemistry. 1983 May 10;22(10):2319–2323. doi: 10.1021/bi00279a003. [DOI] [PubMed] [Google Scholar]
  34. Tedeschi H., Mannella C. A., Bowman C. L. Patch clamping the outer mitochondrial membrane. J Membr Biol. 1987;97(1):21–29. doi: 10.1007/BF01869611. [DOI] [PubMed] [Google Scholar]
  35. Thieffry M., Chich J. F., Goldschmidt D., Henry J. P. Mise en évidence d'un canal ionique de grande conductance dans des fractions sub-cellulaires de la médullo-surrénale. C R Acad Sci III. 1987;305(6):193–197. [PubMed] [Google Scholar]
  36. von EULER U., LISHAJKO F. Improved technique for the fluorimetric estimation of catecholamines. Acta Physiol Scand. 1961 Apr;51:348–355. doi: 10.1111/j.1748-1716.1961.tb02128.x. [DOI] [PubMed] [Google Scholar]

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