Feedback inhibition of L-glutamine D-fructose 6-phosphate amidotransferase by uridine diphosphate N-acetylglucosamine in Neurospora crassa.
The enzyme, l-glutamine d-fructose 6-phosphate amidotransferase (EC 2.6.1.16) of Neurospora crassa, which catalyzes the formation of glucosamine 6-phosphate was shown to be subject to feedback inhibition by uridine diphosphate N-acetyl-d-glucosamine (UDP-GlcNAc). The conclusion is based on the following observations. UDP-GlcNAc, the direct precursor of chitin, did not accumulate in the cell even when its utilization for the synthesis of cell wall chitin was interrupted by the antibiotic polyoxin D, a competitive inhibitor of the chitin synthetase (EC 2.4.1.16). Furthermore, the cellular level of UDP-GlcNAc rose in a short period of time when the amidotransferase was bypassed in vivo by the addition of glucosamine to the growing medium of the fungus. The amidotransferase was purified from N. crassa approximately 85-fold. Kinetic studies showed that UDP-GlcNAc was a potent and specific inhibitor of the amidotransferase, and that it did not alter the Michaelis constant for either l-glutamine or d-fructose 6-phosphate, suggesting that the inhibitor binds at a site on the enzyme distinct from the active site.
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001665:Binding Sites; D002247:Carbon Isotopes; D002473:Cell Wall / Q000378:metabolism; D002686:Chitin / Q000096:biosynthesis; D004586:Electrophoresis; D005246:Feedback; D005944:Glucosamine / Q000378:metabolism / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors; D006600:Hexosephosphates / Q000378:metabolism; D009491:Neurospora / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D010209:Paper; D013053:Spectrophotometry; D000637:Transaminases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D014500:Uracil Nucleotides / Q000378:metabolism / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D002247:Carbon Isotopes; D006600:Hexosephosphates; D014500:Uracil Nucleotides; D002686:Chitin; D005964:Glucosyltransferases; D000637:Transaminases; D005947:Glucose; D005944:Glucosamine