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J. Biochem, 1967, Vol. 62, No. 3 328-338
© 1967 Japanese Biochemical Society

research-article

Studies on Amino-acyl-sRNA Synthetases from Pseudomonas aeruginosa

I. Partial Purification of Leucyl-, Phenylalanyl-, Tyrosyl-, Isoleucyl-, and Methionyl-sRNA Synthetases*

YASUSHI TAKASASHI**, MASAKO SATO-KUZUYA*** and YOSHITO KAZIRO***

From the Department of Biochemistry, Faculty of Medicine, University of Tokyo Bunkyo-ku, Tokyo

A partial purification procedure is described for leucyl-, phenylalanyl-, tyrosyl-, isoleucyl-, and methionyl-sRNA synthetases from Pseudomonas aeruginosa. The purification steps included: sonic disruption of cells, removal of nucleic acids by protamine sulfate treatment, ammonium sulfate fractionation, DEAE-cellulose column chromatography, and hydroxylapatite column chromatography. The specific activities of the purified amino-acyl-sRNA synthetases are(in µmoles 32PP1 incorporated into ATP/hour/mg. protein): 882(leucine), 239(phenylalanine), 282(tyro-sine), 400(isoleucine), and 423(methionine).

The purified preparations of leucyl-, phenylalanyl-, and tyrosyl-sRNA synthetases are essentially free from any other amino-acyl-sRNA synthetase activity. Isoleucyl-, and methionyl-sRNA synthetases are purified in the same fraction throughout the present purification procedure. The two activities are, however, separated by a sucrose gradient centrifugation.

The sedimentation coefficients relative to bovine liver catalase are calculated as 8.9S, 6.8S, 5.8S, 5.4S, and 5.0S, for phenylalanyl-, methionyl-, isoleucyl-, leucyl-, and tyrosyl-sRNA synthetases, respectively.

* Dedicated to Dr. K. Kaziro, Professor Emerituj of the Nippon Medical School, on the occasion of his 70th birthday. One of the authors (Y.K.) appreciates him with filial affection the transmission of biochemical and genetical heritage.

** On leave of absence from the Nippon Kayaku Co., Ltd. Present address: Ohji Pharmaceutical Works, Nippon Kayaku Co., Ltd., Kita-ku Tokyo.

***Present address: Department of Chemistry, Institute of Medical Science (formerly known as the Institute for Infectious Diseases), University of Tokyo. Minato-ku, Tokyo. The reprint request should be addressed to Y. Kaziro.


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