Subunit f of the yeast mitochondrial ATP synthase: topological and functional studies.

Abstract

Modified versions of subunit f were produced by mutagenesis of the ATP17 gene of Saccharomyces cerevisiae. A version of subunit f devoid of the last 28 amino acid residues including the unique transmembranous domain complemented the oxidative phosphorylation of the null mutant. However, a two-fold decrease in the specific ATP synthase activity was measured and attributed to a decrease in the stability of the mutant ATP synthase complex as shown by the low oligomycin-sensitive ATPase activity at alkaline pH. The modification or not by nonpermeant maleimide reagents of cysteine residues introduced at the N and C termini of subunit f indicated a Nin-Cout orientation. From the C terminus of subunit f it was possible to cross-link subunit 4 (also called subunit b), which is another component of the F0 sector and which also displays a short hydrophilic segment exposed to the intermembrane space.