Structure of Outward-Facing PglK and Molecular Dynamics of Lipid-Linked Oligosaccharide Recognition and Translocation

Structure
Camilo PerezKaspar P Locher

Abstract

PglK is a lipid-linked oligosaccharide (LLO) flippase essential for asparagine-linked protein glycosylation in Campylobacter jejuni. Previously we have proposed a non-alternating-access LLO translocation mechanism, where postulated outward-facing states play a primary role. To investigate this unusual mechanistic proposal, we have determined a high-resolution structure of PglK that displays an outward semi-occluded state with the two nucleotide binding domains forming an asymmetric closed dimer with two bound ATPγS molecules. Based on this structure, we performed extensive molecular dynamics simulations to investigate LLO recognition and flipping. Our results suggest that PglK may employ a "substrate-hunting" mechanism to locally increase the LLO concentration and facilitate its jump into the translocation pathway, for which sugars from the LLO head group are essential. We further conclude that the release of LLO to the outside occurs before ATP hydrolysis and is followed by the closing of the periplasmic cavity of PglK.

Citations

Jun 23, 2020·Annual Review of Biochemistry·Christoph Thomas, Robert Tampé
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Jul 25, 2020·Proceedings of the National Academy of Sciences of the United States of America·Chengcheng FanDouglas C Rees
Sep 28, 2020·FEBS Letters·Peter M Jones, Anthony M George
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Feb 27, 2021·Biochemical Society Transactions·Tadaomi Furuta
Jun 10, 2021·Structure·Martin PrescherLutz Schmitt

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