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Proc Natl Acad Sci U S A. 2018 Jun 5;115(23):5962-5967. doi: 10.1073/pnas.1800647115. Epub 2018 May 21.

Structure of an EIIC sugar transporter trapped in an inward-facing conformation.

Author information

1
Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030.
2
Department of Biological Sciences, Lehigh University, Bethlehem, PA 18015.
3
Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX 77030.
4
Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
5
Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX 77030; mzhou@bcm.edu allan.ferreon@bcm.edu woi216@lehigh.edu.
6
Department of Biological Sciences, Lehigh University, Bethlehem, PA 18015; mzhou@bcm.edu allan.ferreon@bcm.edu woi216@lehigh.edu.
7
Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030; mzhou@bcm.edu allan.ferreon@bcm.edu woi216@lehigh.edu.

Abstract

The phosphoenolpyruvate-dependent phosphotransferase system (PTS) transports sugar into bacteria and phosphorylates the sugar for metabolic consumption. The PTS is important for the survival of bacteria and thus a potential target for antibiotics, but its mechanism of sugar uptake and phosphorylation remains unclear. The PTS is composed of multiple proteins, and the membrane-embedded Enzyme IIC (EIIC) component transports sugars across the membrane. Crystal structures of two members of the glucose superfamily of EIICs, bcChbC and bcMalT, were solved in the inward-facing and outward-facing conformations, and the structures suggest that sugar translocation could be achieved by movement of a structured domain that contains the sugar-binding site. However, different conformations have not been captured on the same transporter to allow precise description of the conformational changes. Here we present a crystal structure of bcMalT trapped in an inward-facing conformation by a mercury ion that bridges two strategically placed cysteine residues. The structure allows direct comparison of the outward- and inward-facing conformations and reveals a large rigid-body motion of the sugar-binding domain and other conformational changes that accompany the rigid-body motion. All-atom molecular dynamics simulations show that the inward-facing structure is stable with or without the cross-linking. The conformational changes were further validated by single-molecule Föster resonance energy transfer (smFRET). Combined, these results establish the elevator-type mechanism of transport in the glucose superfamily of EIIC transporters.

KEYWORDS:

bcMalT; double-cysteine cross-linking; elevator-type mechanism of transport; inward-facing conformation; single-molecule FRET

PMID:
29784777
DOI:
10.1073/pnas.1800647115
[Indexed for MEDLINE]
Free PMC Article

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