Synthesis of 5-IASA-UDP-GlcNAc and its use for the photoaffinity labeling of a novel UDP-GlcNAc pyrophosphorylase.
Photoreactivable 5-[3-(p-iodoazidosalicylamide)allyl]-UDP-GlcNAc (5-IASA-UDP-GlcNAc) was synthesized by a four-step procedure and used for photoaffinity labeling of UDP-GlcNAc-dependent enzymes. Upon iodination with 125I, the compound was successfully applied to probe a purified UDP-GlcNAc pyrophosphorylase from pig liver. The enzyme was photoinactivated by the probe in the concentration-dependent manner, and was protected by UDP-GlcNAc and, to a lesser extent, by UTP and UDP-GlcCOOH.
Szumiło
H
H
Department of Inorganic and Analytical Chemistry, Medical School, Lublin, Poland.
Szumiło
T
T
Elbein
A D
AD
D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry; D000818:Animals; D001386:Azides / Q000138:chemical synthesis* / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes; D008099:Liver / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D010777:Photochemistry; D013050:Spectrometry, Fluorescence; D013552:Swine; D014530:Uridine Diphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000737:chemistry
C095318:5-(3-(4-iodoazidosalicylamide)allyl)-UDP-N-acetylglucosamine; D000345:Affinity Labels; D001386:Azides; D007457:Iodine Radioisotopes; D014537:Uridine Diphosphate N-Acetylglucosamine; D014530:Uridine Diphosphate; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase