Cell-Free Amino Acid-Incorporating System from Pseudomonas indigofera

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 C 14 -phenylalanine and C 14 -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 p H optimum of 7.6, had many characteristics of protein synthesis. The process depended upon both “ribosomes” and supernatant fraction from centrifugation at 105,000 × 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. C 14 -phenylalanine uptake was stimulated by sRNA or polyuridylic acid, and together these substances had a synergistic effect upon stimulation. The incorporation of C 14 -phenylalanine into a product which was precipitated by antiserum to crystalline isocitrate lyase was also observed.