Regulation of purine utilization in bacteria. VI. Characterization of hypoxanthine and guanine uptake into isolated membrane vesicles from Salmonella typhimurium

L E Jackman, J Hochstadt
1976 Journal of Bacteriology  
Uptake of hypoxanthine and guanine into isolated membrane vesicles of Salmonella typhimurium TR119 was stimulated by 5'-phosphoribosyl-1'-pyrophosphate (PRPP). For strain proAB47, a mutant that lacks guanine phosphoribosyltransferase, PRPP stimulated uptake of hypoxanthine into membrane vesicles. No PRPP-stimulated uptake of guanine was observed. For strain TR119, guanosine 5'-monophosphate and inosine 5'-monophosphate accumulated intravesicularly when guanine and hypoxanthine, respectively,
more » ... e used with PRPP as transport substrates. For strain proAB47, IMP accumulated intravesicularly with hypoxanthine and PRPP as transport substrates. For strain TR1 19, hypoxanthine also accumulated when PRPP was absent. This free hypoxanthine uptake was completely inhibited by N-ethylmaleimide, but the PRPP-stimulated uptake of hypoxanthine was inhibited only 20% by N-ethylmaleimide. Hypoxanthine and guanine phosphoribosyltransferase activity paralleled uptake activity in both strains. But, when proAB47 vesicles were sonically treated to release the enzymes, a threeto sixfold activation of phosphoribosyltransferase molecules occurred. Since proAB47 vesicles lack the guanine phosphoribosyltransferase gene product and since hypoxanthine effectively competes out the phosphoribosylation of guanine by proAB47 vesicles, it was postulated that the hypoxanthine phosphoribosyltransferase gains specificity for both guanine and hypoxanthine when released from the membrane. A group translocation as the major mechanism for the uptake of guanine and hypoxanthine was proposed. Three basic mechanisms of substrate transport have been identified in bacteria. the facilitated diffusion of glycine exemplifies a nonenergized, non-concentrative carrier-mediated mechanism (17, 18). The concentrative uptake of lactose, proline, and other amino acids in enteric bacteria, coupled to a lactic dehydrogenase-mediated energized membrane state (1, 19, 20, 27) , represents a second transport mechanism. In both of these systems, the substrate without alteration traverses the membrane. The third type of transport reaction, in which covalent change of the substrate occurs during transport, is referred to as group translocation. This mechanism is exemplified by the phosphoenolpyruvate-phosphotransferase system which converts a variety of monosaccharides to their sugar phosphates during the permeation process (22, 23, 24) , and by the adenine phosphoribosyl-transferase system which, in the Present address:
doi:10.1128/jb.126.1.312-326.1976 fatcat:rlbkihfesvcpbhddhthwqusvjm