Structural basis of substrate binding in WsaF, a rhamnosyltransferase from Geobacillus stearothermophilus.

Journal of Molecular Biology
Kerstin SteinerJames H Naismith

Abstract

Carbohydrate polymers are medically and industrially important. The S-layer of many Gram-positive organisms comprises protein and carbohydrate polymers and forms an almost paracrystalline array on the cell surface. Not only is this array important for the bacteria but it has potential application in the manufacture of commercially important polysaccharides and glycoconjugates as well. The S-layer glycoprotein glycan from Geobacillus stearothermophilus NRS 2004/3a is mainly composed of repeating units of three rhamnose sugars linked by alpha-1,3-, alpha-1,2-, and beta-1,2-linkages. The formation of the beta-1,2-linkage is catalysed by the enzyme WsaF. The rational use of this system is hampered by the fact that WsaF and other enzymes in the pathway share very little homology to other enzymes. We report the structural and biochemical characterisation of WsaF, the first such rhamnosyltransferase to be characterised. Structural work was aided by the surface entropy reduction method. The enzyme has two domains, the N-terminal domain, which binds the acceptor (the growing rhamnan chain), and the C-terminal domain, which binds the substrate (dTDP-beta-l-rhamnose). The structure of WsaF bound to dTDP and dTDP-beta-l-rhamnose coupled to...Continue Reading

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Citations

Mar 5, 2010·Glycobiology·Kristof ZarschlerChristina Schäffer
Nov 21, 2013·Glycobiology·David Albesa-JovéMarcelo E Guerin
Sep 28, 2010·International Journal of Microbiology·Robin RistlChristina Schäffer
May 28, 2010·Protein Science : a Publication of the Protein Society·Thomas KowatzJames H Naismith
May 20, 2011·Current Opinion in Biotechnology·Aram ChangJon S Thorson
Aug 28, 2016·FEMS Microbiology Reviews·Christina Schäffer, Paul Messner
Feb 19, 2021·The Biochemical Journal·Ben A WagstaffHelge C Dorfmueller
Apr 6, 2012·Carbohydrate Research·Laura K Greenfield, Chris Whitfield
Dec 15, 2018·Progress in Biophysics and Molecular Biology·Devendra K DhakedLalitha Guruprasad

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Methods Mentioned

BETA
PISA
gel filtration
X-ray

Software Mentioned

PISA Interfaces , Surfaces and Assemblies
HYSS
Phaser
BLAST
MassLynx
PyMOL
MicroCal Origin
AUTOSOLVE
Coot
XDS

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