Journal article
Heme Histidine Ligands within gp91phox Modulate Proton Conduction by the Phagocyte NADPH Oxidase
The Journal of biological chemistry, Vol.276(32), pp.30277-30284
08/10/2001
DOI: 10.1074/jbc.M010438200
Abstract
The membrane subunit of the phagocyte NADPH oxidase, gp91phox, possesses a H+ channel motif formed by membrane-spanning histidines postulated to coordinate the two heme groups forming the redox center of the flavocytochrome. To study the role of heme-binding histidines on proton conduction, we stably expressed the gp91phox cytochrome in human embryonic kidney 293 cells and measured proton currents with the patch clamp technique. Similar to its shorter homologue, NADPH oxidase homologue 1, which is predicted not to bind heme, gp91phox generated voltage-activated, pH-dependent, H+-selective currents that were reversibly blocked by Zn2+. The gp91phox currents, however, activated faster, deactivated more slowly, and were markedly affected by the inhibition of heme synthesis. Upon heme removal, the currents had larger amplitude, activated faster and at lower voltages, and became sensitive to the histidine reagent diethylpyrocarbonate. Mutation of the His-115 residue to leucine abolished both the gp91phoxcharacteristic 558-nm absorbance peak and voltage-activated currents, indicating that His-115 is involved in both heme ligation and proton conduction. These results indicate that the gp91phox proton channel is activated upon release of heme from its His-115 ligand. During activation of the oxidase complex, changes in heme coordination within the cytochrome might increase the mobility of histidine ligands, thereby coupling electron and proton transport.
Details
- Title: Subtitle
- Heme Histidine Ligands within gp91phox Modulate Proton Conduction by the Phagocyte NADPH Oxidase
- Creators
- Andrés Maturana - University of GenevaSerge Arnaudeau - University Hospital of GenevaStephan Ryser - University of GenevaBotond Banfi - University of GenevaJohann Peter Hossle - University Children's Hospital ZurichWerner Schlegel - University of GenevaKarl-Heinz Krause - University of GenevaNicolas Demaurex - University Hospital of Geneva
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.276(32), pp.30277-30284
- DOI
- 10.1074/jbc.M010438200
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 08/10/2001
- Academic Unit
- Anatomy and Cell Biology; Otolaryngology; Internal Medicine
- Record Identifier
- 9984284458702771
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