Synthesis of Saccharomyces cerevisiae Catalase A in vitro

Abstract

MRNA isolated from cells of the yeast S. cerevisiae was translated in the cell-free protein synthesis system from wheat germ. Catalase proteins synthesized were isolated from incubation mixtures by immunoadsorption followed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. On dodecyl sulfate gels, catalase A synthesized by the wheat germ system migrates like catalase T protein synthesized in vivo. Yeast catalase T and A synthesized in vivo are not glycoproteins. Synthesis of the 2 catalase proteins in the wheat germ system and dissimilarity of proteolytic fingerprints of the 2 proteins demonstrate conclusively that catalase T and A are biogenetically unrelated.