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Membrane association of monotopic phosphoglycosyl transferase underpins function

Abstract

Polyprenol phosphate phosphoglycosyl transferases (PGTs) catalyze the first membrane-committed step in assembly of essential glycoconjugates. Currently there is no structure–function information to describe how monotopic PGTs coordinate the reaction between membrane-embedded and soluble substrates. We describe the structure and mode of membrane association of PglC, a PGT from Campylobacter concisus. The structure reveals a unique architecture, provides mechanistic insight and identifies ligand-binding determinants for PglC and the monotopic PGT superfamily.

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Fig. 1: PglC reveals a distinct architecture and topology for monotopic membrane proteins.
Fig. 2: PglC crystallizes in a native conformation.
Fig. 3: The active site structure supports the proposed mechanism of PglC.

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Acknowledgements

We thank K. Rajashankar for assistance with phasing and the staff at NECAT (APS) for facilitating X-ray data collection. Financial support for this work was provided by the National Institutes of Health: R01-GM039334 to B.I., the Predoctoral Training Program in the Biological Sciences (T32-GM007287) to S.E. and the Biomolecular Pharmacology Program Grant (T32-GM008541) to L.C.R. This work is also based upon research conducted at the Northeastern Collaborative Access Team beamlines 24-ID-E and 24-ID-C, which is funded by the National Institute of General Medical Sciences from the National Institutes of Health (P41 GM103403).

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Contributions

L.C.R. crystallized, collected data, determined the structure, refined and analyzed the model of PglC and performed phosphate release kinetics; D.D. optimized expression, designed and made constructs, expressed and purified PglC, carried out lipid analysis and analyzed the structure. S.E. designed and performed SCAM analyses. V.L. designed and purified original constructs for crystallization and A.J.L. obtained the original crystallization conditions. L.C.R., D.D. and S.E. wrote the manuscript. B.I. and K.N.A. conceived the project, designed experiments, assisted with data analysis and interpretation, and critically edited the manuscript.

Corresponding authors

Correspondence to Barbara Imperiali or Karen N. Allen.

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Supplementary Text and Figures

Supplementary Figures 1–11 and Supplementary Tables 1–5

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Dataset 1

Supplementary Dataset 1

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Ray, L.C., Das, D., Entova, S. et al. Membrane association of monotopic phosphoglycosyl transferase underpins function. Nat Chem Biol 14, 538–541 (2018). https://doi.org/10.1038/s41589-018-0054-z

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