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Glucose‐6‐phosphate as a probe for the glucosamine‐6‐phosphate N ‐acetyltransferase Michaelis complex
Author(s) -
Hurtado-Guerrero Ramon,
Raimi Olawale,
Shepherd Sharon,
van Aalten Daan M.F.
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.10.065
Subject(s) - substrate (aquarium) , chemistry , transferase , phosphate , glucosamine , acetylation , nucleophile , acetyltransferase , protonation , transition state analog , enzyme , amine gas treating , stereochemistry , biochemistry , active site , organic chemistry , biology , catalysis , gene , ion , ecology
Glucosamine‐6‐phosphate N ‐acetyltransferase (GNA1) catalyses the N ‐acetylation of d ‐glucosamine‐6‐phosphate (GlcN‐6P), using acetyl‐CoA as an acetyl donor. The product GlcNAc‐6P is an intermediate in the biosynthesis UDP‐GlcNAc. GNA1 is part of the GCN5‐related acetyl transferase family (GNATs), which employ a wide range of acceptor substrates. GNA1 has been genetically validated as an antifungal drug target. Detailed knowledge of the Michaelis complex and trajectory towards the transition state would facilitate rational design of inhibitors of GNA1 and other GNAT enzymes. Using the pseudo‐substrate glucose‐6‐phosphate (Glc‐6P) as a probe with GNA1 crystals, we have trapped the first GNAT (pseudo‐)Michaelis complex, providing direct evidence for the nucleophilic attack of the substrate amine, and giving insight into the protonation of the thiolate leaving group.