Abstract
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. (c) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
| Original language | English |
|---|---|
| Pages (from-to) | 5597-5600 |
| Number of pages | 4 |
| Journal | FEBS Letters |
| Volume | 581 |
| Issue number | 29 |
| DOIs | |
| Publication status | Published - 11 Dec 2007 |
Keywords
- Crystal structure
- Michaelis complex
- Acetyltransferase
- UDP-GlcNAc
- Kinetics
- Inhibitor
- Blastocladiella emersonii
- Hexosamine biosynthesis
- Acetylglucosamine
- Protein