Accelerated Evolution of Cytochrome c in Higher Primates, and Regulation of the Reaction between Cytochrome c and Cytochrome Oxidase by Phosphorylation

by Brand, Sue Ellen; Scharlau, Martha; Geren, Lois; Hendrix, Marissa; Parson, Clayre; Elmendorf, Tyler; Neel, Earl; Pianalto, Kaila; Silva-Nash, Jennifer; Durham, Bill; Millett, Francis

Cytochrome c (Cc) underwent accelerated evolution from the stem of the anthropoid primates to humans. Of the 11 amino acid changes that occurred from horse Cc to human Cc, five were at Cc residues near the binding site of the Cc:CcO complex. Single-point mutants of horse and human Cc were made at each of these positions. The Cc:CcO dissociation constant K-D of the horse mutants decreased in the order: T89E > native horse Cc > V11I Cc > Q12M > D50A > A83V > native human. The largest effect was observed for the mutants at residue 50, where the horse Cc D50A mutant decreased K-D from 28.4 to 11.8 mu M, and the human Cc A50D increased K-D from 4.7 to 15.7 mu M. To investigate the role of Cc phosphorylation in regulating the reaction with CcO, phosphomimetic human Cc mutants were prepared. The Cc T28E, S47E, and Y48E mutants increased the dissociation rate constant k(d), decreased the formation rate constant k(f), and increased the equilibrium dissociation constant K-D of the Cc:CcO complex. These studies indicate that phosphorylation of these residues plays an important role in regulating mitochondrial electron transport and membrane potential Delta psi.

Journal
Cells
Volume
11
Issue
24
Year
2022
URL
https://dx.doi.org/10.3390/cells11244014
ISBN/ISSN
2073-4409
DOI
10.3390/cells11244014