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A coiled-coil domain acts as a molecular ruler to regulate O-antigen chain length in lipopolysaccharide

DOI: 10.1038/nsmb.2935 DOI Help
PMID: 25504321 PMID Help

Authors: Gregor Hagelueken (Centre for Biomolecular Sciences, The University of St. Andrews) , Bradley R Clarke (University of Guelph) , Hexian Huang (University of St Andrews) , Anne Tuukkanen (European Molecular Biology Laboratory, Hamburg Outstation) , Iulia Danciu (European Molecular Biology Laboratory, Hamburg Outstation) , Dmitri I Svergun (European Molecular Biology Laboratory, Hamburg Outstation) , Rohanah Hussain (Diamond Light Source) , Huanting Liu (University of St. Andrews) , Chris Whitfield (University of Guelph) , James Naismith (Center for Biomolecular Sciences, University of St. Andrews)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Structural & Molecular Biology

State: Published (Approved)
Published: December 2014
Diamond Proposal Number(s): 11051 , 12036

Open Access Open Access

Abstract: Long-chain bacterial polysaccharides have important roles in pathogenicity. In Escherichia coli O9a, a model for ABC transporter–dependent polysaccharide assembly, a large extracellular carbohydrate with a narrow size distribution is polymerized from monosaccharides by a complex of two proteins, WbdA (polymerase) and WbdD (terminating protein). Combining crystallography and small-angle X-ray scattering, we found that the C-terminal domain of WbdD contains an extended coiled-coil that physically separates WbdA from the catalytic domain of WbdD. The effects of insertions and deletions in the coiled-coil region were analyzed in vivo, revealing that polymer size is controlled by varying the length of the coiled-coil domain. Thus, the coiled-coil domain of WbdD functions as a molecular ruler that, along with WbdA:WbdD stoichiometry, controls the chain length of a model bacterial polysaccharide.

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

Instruments: B23-Circular Dichroism , I04-Macromolecular Crystallography

Other Facilities: X33, EMBL

Added On: 16/12/2014 12:12

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