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J Bacteriol. 1965 October; 90(4): 978-983
Copyright © 1965 American Society for Microbiology. All Rights Reserved.

Cell-Free Amino Acid-Incorporating System from Pseudomonas indigofera

Department of Chemistry, Washington State University, Pullman, Washington

ABSTRACT

SHIIO, TSURU (Washington State University, Pullman), AND BRUCE A. MCFADDEN. Cell-free amino acid-incorporating system from Pseudomonas indigofera. J. Bacteriol. 90:978–983. 1965.—A cell-free preparation from Pseudomonas indigofera incorporated C¹⁴-phenylalanine and C¹⁴-leucine into a product which was insoluble in hot trichloroacetic acid. The phenylalanine incorporation process, which had a temperature optimum of 30 C and a pH optimum of 7.6, had many characteristics of protein synthesis. The process depended upon both "ribosomes" and supernatant fraction from centrifugation at 105,000 x g. Incorporation required adenosine triphosphate, apparently depended upon guanosine triphosphate, and was inhibited by chloramphenicol, puromycin, actinomycin, ribonuclease, and deoxyribonuclease. Leucine incorporation was also studied and had many similar characteristics. C¹⁴-phenylalanine uptake was stimulated by sRNA or polyuridylic acid, and together these substances had a synergistic effect upon stimulation. The incorporation of C¹⁴-phenylalanine into a product which was precipitated by antiserum to crystalline isocitrate lyase was also observed.