Kj. Ceesay et al., THE RELATIONSHIP BETWEEN THE TRIMETHYLATION OF LYSINE-77 AND CYTOCHROME-C METABOLISM IN SACCHAROMYCES-CEREVISIAE, International Journal of Biochemistry, 26(5), 1994, pp. 721-734
1. Site directed mutations were constructed in the yeast iso-1-cytochr
ome c gene adjacent to the lysine 77 (methylation site) codon. 2. Thes
e mutant genes were then cloned and transformed into the S. cerevisiae
strain B-6642 which contains a deficiency in the iso-1-cytochrome c g
ene. 3. The resulting transformants were screened for cytochrome c pro
duction using gel electrophoresis. 4. Amino acid analysis of the mutat
ed cytochromes c demonstrated varying levels of trimethyllysine format
ion, depending on the nature of the site directed mutation. 5. The res
ulting transformants were then used as tools in order to investigate t
he relationship between trimethyllysine formation and various aspects
of cytochrome c metabolism including protein stability and heme conjug
ation.