Physiological consequences of starvation in Pseudomonas putida: degradation of intracellular protein and loss of activity of the inducible enzymes of L-arginine catabolism.
We investigated the degradation of radioisotopically labeled intracellular protein in starved, intact cells of Pseudomonas putida P2 (ATCC 25571) and the regulation of this process. Intracellular protein isotopically labeled with L-[4,5-3H]leucine during log-phase growth at 30 C is degraded at rates of 1 to 2%/h in log-phase cells and 7 to 9%/h in starved cells. Rifampin, chloramphenicol, and tosyllysine chloromethylketone lower the rate of protein degradation by starved cells. Addition to starved cells of a nutrient upon which the culture is induced for growth rapidly lowers the rate of protein degradation from 7 to 9%/h to less than 1.5%/h. A nutrient that is oxidized but that cannot immediately support growth also lowers the rate of starvation-induced protein degradation. Proteolytic activity of cell extracts requires a divalent metal ion and may be inhibited up to 60% by tosyllysine chloromethylketone or p-toluenesulfonyl fluoride. Rifampin and chloramphenicol have no effect. In contrast to intact cells, extracts of growing or starving cells degrade protein at equivalent rates. We also investigated the stabilities of the inducible transport system and of four inducible intracellular enzymes of L-arginine catabolism. These include: the membrane-associated, L-arginine-specific transport system; L-arginine oxidase (oxidase); alpha-ketoarginine decarboxylase (decarboxylase); gamma-guanidinobutyraldehyde dehydrogenase ( dehydrogenase); and gamma-guanidinobutyrate amidinohydrolase (hydrolase). In starved cells, the rates of loss of activities were: transport and dehydrogenase activities, stable; oxidase and decarboxylase activities, 20 to 30%/h; hydrolase activity, 5 to 8%/h. Chloramphenicol decreases the rate of loss of oxidase, decarboxylase, and hydrolase activity, whereas p-toluenesulfonyl fluoride lowers the rate of loss of decarboxylase but not of oxidase or hydrolase activity. Addition to starved cells of a nutrient for which they are already induced for growth (e.g., malate, a noninducer of arginine catabolic enzymes) decreases the rate of loss of oxidase and decarboxylase activity but not that of the hydrolase.
D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism; D001120:Arginine / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002087:Butyrates; D002262:Carboxy-Lyases / Q000378:metabolism; D002474:Cell-Free System; D002701:Chloramphenicol / Q000494:pharmacology; D006867:Hydrolases / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D012293:Rifampin / Q000494:pharmacology; D013237:Stereoisomerism; D014105:Tosyl Compounds / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
D001426:Bacterial Proteins; D002087:Butyrates; D008293:Malates; D014105:Tosyl Compounds; D014107:Tosyllysine Chloromethyl Ketone; D002701:Chloramphenicol; D000641:Ammonia; D001120:Arginine; D010105:Oxygenases; D000445:Aldehyde Oxidoreductases; D006867:Hydrolases; D002262:Carboxy-Lyases; D012293:Rifampin