Release of rhodanese from Pseudomonas aeruginosa by cold shock and its localization within the cell.
Whole cells of Pseudomonas aeruginosa possess rhodanese activity. The enzyme can be released by rapidly resuspending the cells in cold Tris--HCl buffer. Approximately 95% of the rhodanese activity is released by cold shock. Release of the enzyme can be inhibited either by preincubating the cells with Mg2+ or by incorporating Mg2+ into the shocking buffer. The effect of Mg2+ can be reversed by washing the cells twice with buffer prior to cold shock. While rhodanese can be released from P. aeruginosa by cold shock, lactic dehydrogenase, a cytoplasmic enzyme, remains within the cell. Diazo-7-amino-1,3-napthalenedisulfonic acid, a compound which does not penetrate the cytoplasmic membrane, completely inactivated rhodanese and alkaline phosphatase, a periplasmic enzyme, whereas lactic dehydrogenase retained its full activity. These data suggest that rhodanese in P. aeruginosa, like alkaline phosphatase, is located distal to the cytoplasmic membrane in the periplasmic space. Electron micrographs also show that portions of the lipopolysaccharide outer membrane are shed from the cell during cold shock, while cells preincubated with Mg2+ did not release segments of their outer membrane.
D000469:Alkaline Phosphatase / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D003080:Cold Temperature; D003593:Cytoplasm / Q000201:enzymology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D009282:Naphthalenesulfonates / Q000494:pharmacology; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000648:ultrastructure; D013466:Sulfurtransferases / Q000302:isolation & purification*; D013884:Thiosulfate Sulfurtransferase / Q000302:isolation & purification* / Q000378:metabolism; D014871:Water Microbiology
D009282:Naphthalenesulfonates; D007770:L-Lactate Dehydrogenase; D013466:Sulfurtransferases; D013884:Thiosulfate Sulfurtransferase; D000469:Alkaline Phosphatase; D008274:Magnesium