RRC ID 54390
Author Kato HE, Inoue K, Abe-Yoshizumi R, Kato Y, Ono H, Konno M, Hososhima S, Ishizuka T, Hoque MR, Kunitomo H, Ito J, Yoshizawa S, Yamashita K, Takemoto M, Nishizawa T, Taniguchi R, Kogure K, Maturana AD, Iino Y, Yawo H, Ishitani R, Kandori H, Nureki O.
Title Structural basis for Na(+) transport mechanism by a light-driven Na(+) pump.
Journal Nature
Abstract Krokinobacter eikastus rhodopsin 2 (KR2) is the first light-driven Na(+) pump discovered, and is viewed as a potential next-generation optogenetics tool. Since the positively charged Schiff base proton, located within the ion-conducting pathway of all light-driven ion pumps, was thought to prohibit the transport of a non-proton cation, the discovery of KR2 raised the question of how it achieves Na(+) transport. Here we present crystal structures of KR2 under neutral and acidic conditions, which represent the resting and M-like intermediate states, respectively. Structural and spectroscopic analyses revealed the gating mechanism, whereby the flipping of Asp116 sequesters the Schiff base proton from the conducting pathway to facilitate Na(+) transport. Together with the structure-based engineering of the first light-driven K(+) pumps, electrophysiological assays in mammalian neurons and behavioural assays in a nematode, our studies reveal the molecular basis for light-driven non-proton cation pumps and thus provide a framework that may advance the development of next-generation optogenetics.
Volume 521(7550)
Pages 48-53
Published 2015-5-7
DOI 10.1038/nature14322
PII nature14322
PMID 25849775
MeSH Binding Sites Crystallography, X-Ray Flavobacteriaceae / chemistry* Hydrogen-Ion Concentration Ion Pumps / chemistry* Ion Pumps / genetics Ion Pumps / metabolism Ion Pumps / radiation effects* Ion Transport / genetics Ion Transport / radiation effects Light* Models, Biological Models, Molecular Mutagenesis / genetics Optogenetics Potassium / metabolism Protein Conformation Protein Engineering Retinaldehyde / chemistry Retinaldehyde / metabolism Rhodopsin / chemistry* Rhodopsin / genetics Rhodopsin / metabolism Rhodopsin / radiation effects* Schiff Bases Sodium / metabolism* Structure-Activity Relationship
IF 42.779
Times Cited 106
DNA material pCMV-KR2-3.0-EYFP (RDB16777) pCaMKIIa-KR2-3.0-EYFP-WPRE (RDB16778)