1NVK

T4 phage BGT in complex with UDP and a Mn2+ ion at 1.8 A resolution

Summary for 1NVK

• Entry DOI: 10.2210/pdb1nvk/pdb
• Related: 1JG7 1JIU 1JIX
• Descriptor: DNA beta-glucosyltransferase, MANGANESE (II) ION, URIDINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: glycosyltransferase, gt-b, mn, transferase
• Biological source: Enterobacteria phage T4
• Total number of polymer chains: 1
• Total formula weight: 41271.07

Authors

Lariviere, L.,Kurzeck, J.,Gueguen-Chaignon, V.,Rueger, W.,Morera, S. (deposition date: 2003-02-04, release date: 2003-09-09, Last modification date: 2024-02-14)

Primary citation

Lariviere, L.,Gueguen-Chaignon, V.,Morera, S.
Crystal structures of the T4 phage beta-glucosyltransferase and the D100A mutant in complex with UDP-glucose: glucose binding and identification of the catalytic base for a direct displacement mechanism
J.Mol.Biol., 330:1077-1086, 2003

PubMed Abstract: T4 phage beta-glucosyltransferase (BGT) is an inverting glycosyltransferase (GT) that transfers glucose from uridine diphospho-glucose (UDP-glucose) to an acceptor modified DNA. BGT belongs to the GT-B structural superfamily, represented, so far, by five different inverting or retaining GT families. Here, we report three high-resolution X-ray structures of BGT and a point mutant solved in the presence of UDP-glucose. The two co-crystal structures of the D100A mutant show that, unlike the wild-type enzyme, this mutation prevents glucose hydrolysis. This strongly indicates that Asp100 is the catalytic base. We obtained the wild-type BGT-UDP-glucose complex by soaking substrate-free BGT crystals. Comparison with a previous structure of BGT solved in the presence of the donor product UDP and an acceptor analogue provides the first model of an inverting GT-B enzyme in which both the donor and acceptor substrates are bound to the active site. The structural analyses support the in-line displacement reaction mechanism previously proposed, locate residues involved in donor substrate specificity and identify the catalytic base.

PubMed: 12860129
DOI: 10.1016/S0022-2836(03)00635-1

Experimental method

X-RAY DIFFRACTION (1.8 Å)