Properties of allosteric nicotinamide mononucleotide glycohydrolase from Azotobacter vinelandii: activation and inhibition.
In order to clarify the regulation mechanism of NMN glycohydrolase, a number of purine and pyrimidine nucleotides were tested as activators for the enzyme. Among naturally occurring nucleotides, pppGpp was shown to be the most potent activator (KA = 0.0087 mM). The effectiveness of these nucleotides estimated from Vmax/KA was in the order: pppGpp greater than ppGpp greater than pppppG greater than ppppG greater than pGpp greater than GTP greater than or equal to 2'-GMP greater than or equal to GppppG greater than dGTP greater than ITP greater than GDP = 2'-O-methylGTP. XTP, UTP, dTTP gave no effect. In contrast, ATP, CTP, and 7-methylGTP were inhibitory. A comparison of the data indicated that activation of the enzyme is specific for the base structure, guanine, and that effectiveness of nucleotides as activators results from the negatively charged phosphate moiety. Consistent with these results, inorganic polyphosphates (tri-, tetra-, and pentaphosphate) and 5-phosphoribose 1-pyrophosphate were shown to function as inhibitors of the enzyme. The inhibition mechanism was shown to be competitive with GTP. However, apparent Ki values for these inhibitors were dependent on the activator concentration. Furthermore, a shift of cooperativity type and progressive increase of positive cooperativity concomitant with increase of PRPP concentration were observed. These results indicate that this enzyme may be allosteric and an important regulatory component of pyridine nucleotide cycle metabolism.
D000494:Allosteric Regulation; D001395:Azotobacter / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D006150:Guanine Nucleotides / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010710:Phosphates / Q000494:pharmacology; D011122:Polyphosphates / Q000494:pharmacology
D006150:Guanine Nucleotides; D010710:Phosphates; D011122:Polyphosphates; D006160:Guanosine Triphosphate; D009699:N-Glycosyl Hydrolases; C021881:NMN nucleosidase