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

Gregor Hageluken; Bradley R. Clarke; Hexian Huang; Anne Tuukkanen; Iulia Danciu; Dmitri I. Svergun; Rohanah Hussain; Huanting Liu; Chris Whitfield; James H. Naismith

doi:10.1038/nsmb.2935

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

Gregor Hageluken, Bradley R. Clarke, Hexian Huang, Anne Tuukkanen, Iulia Danciu, Dmitri I. Svergun, Rohanah Hussain, Huanting Liu, Chris Whitfield, James H. Naismith^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

54 Citations (Scopus)

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.

Original language	English
Pages (from-to)	50-56
Number of pages	7
Journal	Nature Structural and Molecular Biology
Volume	22
Issue number	1
Early online date	15 Dec 2014
DOIs	https://doi.org/10.1038/nsmb.2935
Publication status	Published - Jan 2015

Keywords

Escherichia-coli O9A
Protein secondary structure
Circular-dichroism spectra
Klebsiella-pneumoniae
Polysaccharide export
Bifunctional kinase
Injectisome needle
Flagellar hook
Biosynthesis
Transporter

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10.1038/nsmb.2935

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@article{fe62b6f77d9d4e34aa7939e4b2bb9f19,

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

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.",

keywords = "Escherichia-coli O9A, Protein secondary structure, Circular-dichroism spectra, Klebsiella-pneumoniae, Polysaccharide export, Bifunctional kinase, Injectisome needle, Flagellar hook, Biosynthesis, Transporter",

author = "Gregor Hageluken and Clarke, {Bradley R.} and Hexian Huang and Anne Tuukkanen and Iulia Danciu and Svergun, {Dmitri I.} and Rohanah Hussain and Huanting Liu and Chris Whitfield and Naismith, {James H.}",

note = "This work was supported by Wellcome Trust grant 081862 (J.H.N. and C.W.), Senior Investigator Award WT100209MA (J.H.N.), the Natural Sciences, Engineering Research Council of Canada (C.W.), and the German Federal Ministry of Education and Research (BMBF) project BioSCAT (contract no: 05K12YE1 (A.T. and D.I.S.). J.H.N. is a Royal Society Wolfson Merit Award Holder and C.W. is a recipient of a Canada Research Chair.",

year = "2015",

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doi = "10.1038/nsmb.2935",

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pages = "50--56",

journal = "Nature Structural and Molecular Biology",

issn = "1545-9993",

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TY - JOUR

T1 - A coiled-coil domain acts as a molecular ruler to regulate O-antigen chain length in lipopolysaccharide

AU - Hageluken, Gregor

AU - Clarke, Bradley R.

AU - Huang, Hexian

AU - Tuukkanen, Anne

AU - Danciu, Iulia

AU - Svergun, Dmitri I.

AU - Hussain, Rohanah

AU - Liu, Huanting

AU - Whitfield, Chris

AU - Naismith, James H.

N1 - This work was supported by Wellcome Trust grant 081862 (J.H.N. and C.W.), Senior Investigator Award WT100209MA (J.H.N.), the Natural Sciences, Engineering Research Council of Canada (C.W.), and the German Federal Ministry of Education and Research (BMBF) project BioSCAT (contract no: 05K12YE1 (A.T. and D.I.S.). J.H.N. is a Royal Society Wolfson Merit Award Holder and C.W. is a recipient of a Canada Research Chair.

PY - 2015/1

Y1 - 2015/1

N2 - 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.

AB - 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.

KW - Escherichia-coli O9A

KW - Protein secondary structure

KW - Circular-dichroism spectra

KW - Klebsiella-pneumoniae

KW - Polysaccharide export

KW - Bifunctional kinase

KW - Injectisome needle

KW - Flagellar hook

KW - Biosynthesis

KW - Transporter

UR - https://www.nature.com/articles/nsmb.2935#Sec16

U2 - 10.1038/nsmb.2935

DO - 10.1038/nsmb.2935

M3 - Article

SN - 1545-9993

VL - 22

SP - 50

EP - 56

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 1

ER -

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

Abstract

Keywords

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Carb Export: Bacteria Ref: 081862/Z/06/Z: Carbohydrate export in bacteria

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

Abstract

Keywords

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Carb Export: Bacteria Ref: 081862/Z/06/Z: Carbohydrate export in bacteria

Cite this