Insights into SusCD-mediated glycan import by a prominent gut symbiont

Declan A. Gray, Joshua B. R. White, Abraham O. Oluwole, Parthasarathi Rath, Amy J. Glenwright, Adam Mazur, Michael Zahn, Arnaud Baslé, Carl Morland, Sasha L. Evans, Alan Cartmell, Carol V. Robinson, Sebastian Hiller, Neil A. Ranson, David N. Bolam () and Bert van den Berg ()
Additional contact information
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 Baslé: 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

Nature Communications, 2021, vol. 12, issue 1, 1-14

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-020-20285-y

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