Absence of proton channels in COS-7 cells expressing functional NADPH oxidase components

Deri Morgan, Vladimir V. Cherny, Marianne O. Price, Mary C. Dinauer, Thomas E. Decoursey

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is an enzyme of phagocytes that produces bactericidal superoxide anion (O2-) via an electrogenic process. Proton efflux compensates for the charge movement across the cell membrane. The proton channel responsible for the H+ efflux was thought to be contained within the gp91phox subunit of NADPH oxidase, but recent data do not support this idea (DeCoursey, T.E., V.V. Cherny, D. Morgan, B.Z. Katz, and M.C. Dinauer. 2001. J. Biol. Chem. 276:36063-36066). In this study, we investigated electrophysiological properties and superoxide production of COS-7 cells transfected with all NADPH oxidase components required for enzyme function (COSphox). The 7D5 antibody, which detects an extracellular epitope of the gp91phox protein, labeled 96-98% of COSphox cells. NADPH oxidase was functional because COSphox (but not COSWT) cells stimulated by phorbol myristate acetate (PMA) or arachidonic acid (AA) produced superoxide anion. No proton currents were detected in either wild-type COS-7 cells (COSWT) or COSphox cells studied at pHo 7.0 and pHi 5.5 or 7.0. Anion currents that decayed at voltages positive to 40 mV were the only currents observed. PMA or AA did not elicit detectable H+ current in COSWT or COSphox cells. Therefore, gp91phox does not function as a proton channel in unstimulated cells or in activated cells with a demonstrably functional oxidase.

Original languageEnglish
Pages (from-to)571-580
Number of pages10
JournalJournal of General Physiology
Volume119
Issue number6
DOIs
StatePublished - Jun 20 2002
Externally publishedYes

Keywords

  • H channels
  • Phagocytes
  • Respiratory burst
  • Superoxide
  • gp91

Fingerprint Dive into the research topics of 'Absence of proton channels in COS-7 cells expressing functional NADPH oxidase components'. Together they form a unique fingerprint.

  • Cite this