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Proc Natl Acad Sci U S A. 2014 Feb 4;111(5):1784-8. doi: 10.1073/pnas.1324141111. Epub 2014 Jan 22.

Structure of sugar-bound LacY.

Author information

1
Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94158.

Abstract

Here we describe the X-ray crystal structure of a double-Trp mutant (Gly46→Trp/Gly262→Trp) of the lactose permease of Escherichia coli (LacY) with a bound, high-affinity lactose analog. Although thought to be arrested in an open-outward conformation, the structure is almost occluded and is partially open to the periplasmic side; the cytoplasmic side is tightly sealed. Surprisingly, the opening on the periplasmic side is sufficiently narrow that sugar cannot get in or out of the binding site. Clearly defined density for a bound sugar is observed at the apex of the almost occluded cavity in the middle of the protein, and the side chains shown to ligate the galactopyranoside strongly confirm more than two decades of biochemical and spectroscopic findings. Comparison of the current structure with a previous structure of LacY with a covalently bound inactivator suggests that the galactopyranoside must be fully ligated to induce an occluded conformation. We conclude that protonated LacY binds D-galactopyranosides specifically, inducing an occluded state that can open to either side of the membrane.

KEYWORDS:

X-ray structure; conformational change; induced fit; membrane protein; transport

PMID:
24453216
PMCID:
PMC3918835
DOI:
10.1073/pnas.1324141111
[Indexed for MEDLINE]
Free PMC Article

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