Proton exit from the heme–copper oxidase of Escherichia coli
- Helsinki Bioenergetics Group, Department of Medical Chemistry, Institute of Biomedical Sciences and Biocentrum Helsinki, P.O. Box 8, 00014 University of Helsinki, Helsinki, Finland
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Communicated by Helmut Beinert, University of Wisconsin, Madison, WI (received for review August 2, 1998)
Abstract
Pathways of proton entry have been identified in the proton-translocating heme–copper oxidases, but the proton exit pathway is unknown. Here we report experiments with cytochrome bo 3 in Escherichia coli cells that may identify the beginning of the exit pathway. Systematic mutations of arginines 438 and 439 (R481 and R482 in the E. coli enzyme), numbering as in cytochrome aa 3 from bovine heart mitochondria, which interact with the ring D propionates of the two heme groups, reveal that the D propionate of the oxygen-binding heme is involved in proton pumping; its anionic form must be stabilized in order for proton translocation to occur. This may locate the beginning of the pathway by which pumped protons exit from the enzyme structure.
