Toggle Main Menu Toggle Search

Open Access padlockePrints

Substrate and Metal Ion Promiscuity in Mannosylglycerate Synthase

Lookup NU author(s): Dr Morten Nielsen, Dr Louise Tailford, Dr James Flint, Dr Claire Dumon, Professor Harry Gilbert

Downloads

Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Abstract

The enzymatic transfer of the sugar mannose from activated sugar donors is central to the synthesis of a wide range of biologically significant polysaccharides and glycoconjugates. In addition to their importance in cellular biology, mannosyltransferases also provide model systems with which to study catalytic mechanisms of glycosyl transfer. Mannosylglycerate synthase (MGS) catalyzes the synthesis of alpha-mannosyl-D-glycerate using GDP-mannose as the preferred donor species, a reaction that occurs with a net retention of anomeric configuration. Past work has shown that the Rhodothermus marinus MGS, classified as a GT78 glycosyltransferase, displays a GT-A fold and performs catalysis in a metal ion-dependent manner. MGS shows very unusual metal ion dependences with Mg2+ and Ca2+ and, to a lesser extent, Mn2+, Ni2+, and Co2+, thus facilitating catalysis. Here, we probe these dependences through kinetic and calorimetric analyses of wild-type and site-directed variants of the enzyme. Mutation of residues that interact with the guanine base of GDP are correlated with a higher k(cat) value, whereas substitution of His-217, a key component of the metal coordination site, results in a change in metal specificity to Mn2+. Structural analyses of MGS complexes not only provide insight into metal coordination but also how lactate can function as an alternative acceptor to glycerate. These studies highlight the role of flexible loops in the active center and the subsequent coordination of the divalent metal ion as key factors in MGS catalysis and metal ion dependence. Furthermore, Tyr-220, located on a flexible loop whose conformation is likely influenced by metal binding, also plays a critical role in substrate binding.


Publication metadata

Author(s): Nielsen MM, Suits MDL, Yang M, Barry CS, Martinez-Fleites C, Tailford LE, Flint JE, Dumon C, Davis BG, Gilbert HJ, Davies GJ

Publication type: Article

Publication status: Published

Journal: Journal of Biological Chemistry

Year: 2011

Volume: 286

Issue: 17

Pages: 15155-15164

Print publication date: 02/02/2011

ISSN (print): 0021-9258

ISSN (electronic): 1083-351X

Publisher: American Society for Biochemistry and Molecular Biology, Inc.

URL: http://dx.doi.org/10.1074/jbc.M110.199844

DOI: 10.1074/jbc.M110.199844


Altmetrics

Altmetrics provided by Altmetric


Actions

Find at Newcastle University icon    Link to this publication


Share