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Title: Structure of archaerhodopsin-2 at 1.8 Å resolution

Journal Article · · Acta Crystallographica. Section D: Biological Crystallography

The same hexagonal array of archaerhodpsin-2 trimers as observed in the native membrane is reconstituted in a three-dimensional crystal prepared by the membrane fusion method. In this crystal, a pair of conserved glutamate residues in the proton-release pathway is maintained by a low-barrier hydrogen bond. Archaerhodopsin-2 (aR2), the sole protein found in the claret membrane of Halorubrum sp. Aus-2, functions as a light-driven proton pump. In this study, structural analysis of aR2 was performed using a novel three-dimensional crystal prepared by the successive fusion of claret membranes. The crystal is made up of stacked membranes, in each of which aR2 trimers are arranged on a hexagonal lattice. This lattice structure resembles that found in the purple membrane of H. salinarum, except that lipid molecules trapped within the trimeric structure are not distributed with perfect threefold symmetry. Nonetheless, diffraction data at 1.8 Å resolution provide accurate structural information about functionally important residues. It is shown that two glutamates in the proton-release channel form a paired structure that is maintained by a low-barrier hydrogen bond. Although the structure of the proton-release pathway is highly conserved among proton-pumping archaeal rhodopsins, aR2 possesses the following peculiar structural features: (i) the motional freedom of the tryptophan residue that makes contact with the C13 methyl group of retinal is restricted, affecting the formation/decay kinetics of the L state, and (ii) the N-terminal polypeptide folds into an Ω-loop, which may play a role in organizing the higher-order structure.

OSTI ID:
22347758
Journal Information:
Acta Crystallographica. Section D: Biological Crystallography, Vol. 70, Issue Pt 10; Other Information: PMCID: PMC4188009; PMID: 25286853; PUBLISHER-ID: mv5109; OAI: oai:pubmedcentral.nih.gov:4188009; Copyright (c) Kouyama et al. 2014; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0907-4449
Country of Publication:
Denmark
Language:
English