1OI6

Structure determination of the TMP-complex of EvaD

Summary for 1OI6

• Entry DOI: 10.2210/pdb1oi6/pdb
• Related: 1OFN
• Descriptor: PCZA361.16, GLYCEROL, THYMIDINE-5'-PHOSPHATE, ... (4 entities in total)
• Functional Keywords: epimerase, vancomycin group antibiotic, evad, isomerase
• Biological source: AMYCOLATOPSIS ORIENTALIS (STREPTOMYCES)
• Total number of polymer chains: 2
• Total formula weight: 45852.00

Authors

Merkel, A.B.,Naismith, J.H. (deposition date: 2003-06-09, release date: 2004-06-03, Last modification date: 2023-12-13)

Primary citation

Merkel, A.B.,Major, L.L.,Errey, J.C.,Burkart, M.D.,Field, R.A.,Walsh, C.T.,Naismith, J.H.
The Position of a Key Tyrosine in Dtdp-4-Keto-6-Deoxy-D-Glucose-5-Epimerase (Evad) Alters the Substrate Profile for This Rmlc-Like Enzyme
J.Biol.Chem., 279:32684-, 2004

PubMed Abstract: Vancomycin, the last line of defense antibiotic, depends upon the attachment of the carbohydrate vancosamine to an aglycone skeleton for antibacterial activity. Vancomycin is a naturally occurring secondary metabolite that can be produced by bacterial fermentation. To combat emerging resistance, it has been proposed to genetically engineer bacteria to produce analogues of vancomycin. This requires a detailed understanding of the biochemical steps in the synthesis of vancomycin. Here we report the 1.4 A structure and biochemical characterization of EvaD, an RmlC-like protein that is required for the C-5' epimerization during synthesis of dTDP-epivancosamine. EvaD, although clearly belonging to the RmlC class of enzymes, displays very low activity in the archetypal RmlC reaction (double epimerization of dTDP-6-deoxy-4-keto-D-glucose at C-3' and C-5'). The high resolution structure of EvaD compared with the structures of authentic RmlC enzymes indicates that a subtle change in the enzyme active site repositions a key catalytic Tyr residue. A mutant designed to re-establish the normal position of the Tyr increases the RmlC-like activity of EvaD.

PubMed: 15159413
DOI: 10.1074/JBC.M404091200

Experimental method

X-RAY DIFFRACTION (1.4 Å)