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Insights into SusCD-mediated glycan import by a prominent gut symbiont
DOI:
10.1038/s41467-020-20285-y
Authors:
Declan A.
Gray
(Newcastle University)
,
Joshua B. R.
White
(University of Leeds)
,
Abraham O.
Oluwole
(University of Oxford)
,
Parthasarathi
Rath
(University of Basel)
,
Amy J.
Glenwright
(Newcastle University)
,
Adam
Mazur
(University of Basel)
,
Michael
Zahn
(University of Basel)
,
Arnaud
Basle
(Newcastle University)
,
Carl
Morland
(Newcastle University)
,
Sasha L.
Evans
(University of Leeds)
,
Alan
Cartmell
(University of Liverpool)
,
Carol V.
Robinson
(University of Oxford)
,
Sebastian
Hiller
(University of Basel)
,
Neil A.
Ranson
(University of Leeds)
,
David N.
Bolam
(Newcastle University)
,
Bert
Van Den Berg
(Newcastle University)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Nature Communications
, VOL 12
State:
Published (Approved)
Published:
January 2021
Diamond Proposal Number(s):
13587
,
18598

Abstract: In Bacteroidetes, one of the dominant phyla of the mammalian gut, active uptake of large nutrients across the outer membrane is mediated by SusCD protein complexes via a “pedal bin” transport mechanism. However, many features of SusCD function in glycan uptake remain unclear, including ligand binding, the role of the SusD lid and the size limit for substrate transport. Here we characterise the β2,6 fructo-oligosaccharide (FOS) importing SusCD from Bacteroides thetaiotaomicron (Bt1762-Bt1763) to shed light on SusCD function. Co-crystal structures reveal residues involved in glycan recognition and suggest that the large binding cavity can accommodate several substrate molecules, each up to ~2.5 kDa in size, a finding supported by native mass spectrometry and isothermal titration calorimetry. Mutational studies in vivo provide functional insights into the key structural features of the SusCD apparatus and cryo-EM of the intact dimeric SusCD complex reveals several distinct states of the transporter, directly visualising the dynamics of the pedal bin transport mechanism.
Journal Keywords: Bacteriology; Cryoelectron microscopy; Transporters; X-ray crystallography
Diamond Keywords: Bacteria; Gut Microbiota
Subject Areas:
Biology and Bio-materials,
Chemistry
Instruments:
I03-Macromolecular Crystallography
,
I04-1-Macromolecular Crystallography (fixed wavelength)
,
I04-Macromolecular Crystallography
Added On:
13/01/2021 10:31
Documents:
s41467-020-20285-y.pdf
Discipline Tags:
Health & Wellbeing
Biochemistry
Chemistry
Structural biology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)