Structure of a Streptomyces maltosyltransferase GlgE: a homologue of a genetically validated anti-tuberculosis target

DOI: 10.1074/jbc.M111.279315
PMID: 21914799

Authors: Karl Syson (John Innes Centre) , Clare E. M. Stevenson (John Innes Centre) , Martin Rejzek (John Innes Centre) , Shirley A. Fairhurst (John Innes Centre) , Alap Nair (John Innes Centre) , Celia J. Bruton (John Innes Centre) , Robert A. Field (John Innes Centre) , Keith F. Chater (John Innes Centre) , Stephen Bornemann (John Innes Centre) , David M. Lawson (John Innes Centre)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Biological Chemistry

State: Published (Approved)
Published: September 2011
Diamond Proposal Number(s): 1219

Abstract: GlgE is a recently identified (1→4)-α-D-glucan:phosphate α-D-maltosyltransferase involved in α-glucan biosynthesis in bacteria and is a genetically validated anti-tuberculosis drug target. It is a member of the GH13_3 CAZy sub-family for which no structures were previously known. We have solved the structure of GlgE isoform I from Streptomyces coelicolor and shown that this enzyme has the same catalytic and very similar kinetic properties to GlgE from Mycobacterium tuberculosis. The S. coelicolor enzyme forms a homodimer with each subunit comprising five domains including a core catalytic α-amylase-type domain A with a (β/α)8 fold. This domain is elaborated with domain B and two inserts that are specifically configured to define a well conserved donor pocket capable of binding maltose. Domain A, together with domain N from the neighbouring subunit, forms a hydrophobic patch that is close to the maltose binding site and capable of binding cyclodextrins. Cyclodextrins competitively inhibit the binding of maltooligosaccharides to the S. coelicolor enzyme, showing that the hydrophobic patch overlaps with the acceptor binding site. This patch is incompletely conserved in the M. tuberculosis enzyme such that cyclodextrins do not inhibit this enzyme, despite acceptor length specificity being conserved. The crystal structure reveals two further domains, C and S, the latter being a helix bundle not previously reported in GH13 members. The structure provides a framework for understanding how GlgE functions and will help guide the development of inhibitors with therapeutic potential.

Journal Keywords: Binding; Catalysis; Glucosyltransferases; Glycoside; Kinetics; Maltose; Models; Chemical; Models; Molecular; Mycobacterium; Phosphorylation; Protein; Spectrometry; Mass; Matrix-Assisted; Streptomyces

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I02-Macromolecular Crystallography , I03-Macromolecular Crystallography , I04-Macromolecular Crystallography

Added On: 20/09/2011 17:05

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