yeast aminoacyl tRNA synthetases. methodology for their complete or partial purification and comparison of their relative activities under various extraction conditions

• Published in: Biochimie Vol. 59; no. 5/6; pp. 453 - 462
• Main Authors: Kern, D, Dietrich, A, Fasiolo, F, Renaud, M, Giege, R, Ebel, J.P
• Format: Journal Article
• Language: English
• Published: France 1977
• Subjects: Amino Acyl-tRNA Synthetases - isolation & purification; Phenylalanine-tRNA Ligase - isolation & purification; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Valine-tRNA Ligase - isolation & purification
• ISSN: 0300-9084; 1638-6183
• DOI: 10.1016/S0300-9084(77)80050-3

Several fractionation steps are described which can be applied to the partial purification of the 20 aminoacyl-tRNA synthetases from commercial baker's yeast. Comparative experiments performed in the presence or absence of protease inhibitors revealed that some enzymes prepared in the presence of the inhibitor exhibit much higher specific activities than the proteins extracted in the absence of the inhibitor. The methodology reported can be used for the simultaneous preparation of several pure aminoacyl-tRNA synthetases. As examples, the large scale purification of phenylalanyl-and valyl-tRNA synthetases are described.