Sequence Requirement for Trimethylation of Yeast Cytochromec

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Abstract
Lysine 72 (using the vertebrate numbering system) is trimethylated in cytochromes c from fungi and plants but not from higher animals. We have investigated the characteristics of an amino acid sequence required for trimethylation of lysine 72 by examining 21 altered iso-1-cytochromes c from Saccharomyces cerevisiae having single replacements in the region encompassing residues 67 through 77. These results indicated that tyrosine 74 is critical for trimethylation of lysine 72, whereas replacements at other positions did not produce significant diminutions. Various replacements of tyrosine 74 resulted in different levels of inhibition, with the Y74F replacement causing no significant reduction, and the Y74E and Y74K replacements completely or almost completely preventing trimethylation of lysine 72. However, other similarly spaced lysine and tyrosine residues at other sites in the protein did not result in trimethylation of the lysine residue. Thus, a properly situated aromatic residue, determined by the overall conformation of apocytochrome c in the vicinity of lysine 72, appears to be essential for trimethylation.