title: Asymmetric Functional Conversion of Eubacterial Light-driven Ion Pumps.
abstract: In addition to the well-known light-driven outward proton pumps, novel ion-pumping rhodopsins functioning as outward Na(+) and inward Cl(-) pumps have been recently found in eubacteria. They convert light energy into transmembrane electrochemical potential difference, similar to the prototypical archaeal H(+) pump bacteriorhodopsin (BR) and Cl(-) pump halorhodopsin (HR). The H(+), Na(+), and Cl(-) pumps possess the conserved respective DTE, NDQ, and NTQ motifs in the helix C, which likely serve as their functional determinants. To verify this hypothesis, we attempted functional interconversion between selected pumps from each category by mutagenesis. Introduction of the proton-pumping motif resulted in successful Na(+) → H(+) functional conversion. Introduction of the respective characteristic motifs with several additional mutations leads to successful Na(+) → Cl(-) and Cl(-) → H(+) functional conversions, whereas remaining conversions (H(+) → Na(+), H(+) → Cl(-), Cl(-) → Na(+)) were unsuccessful when mutagenesis of 4-6 residues was used. Phylogenetic analysis suggests that a H(+) pump is the common ancestor of all of these rhodopsins, from which Cl(-) pumps emerged followed by Na(+) pumps. We propose that successful functional conversions of these ion pumps are achieved exclusively when mutagenesis reverses the evolutionary amino acid sequence changes. Dependence of the observed functional conversions on the direction of evolution strongly suggests that the essential structural mechanism of an ancestral function is retained even after the gain of a new function during natural evolution, which can be evoked by a few mutations. By contrast, the gain of a new function needs accumulation of multiple mutations, which may not be easily reproduced by limited mutagenesis in vitro.
authors.affiliation: From the Department of Frontier Materials and OptoBioTechnology Research Center, Nagoya Institute of Technology, Showa-ku, Nagoya 466-8555, Japan and PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Inoue
authors.affiliation: From the Department of Frontier Materials and.
authors.forename: Yurika
authors.initials: Y
authors.lastname: Nomura
authors.affiliation: From the Department of Frontier Materials and OptoBioTechnology Research Center, Nagoya Institute of Technology, Showa-ku, Nagoya 466-8555, Japan and kandori@nitech.ac.jp.
authors.forename: Hideki
authors.initials: H
authors.lastname: Kandori
mesh_terms: D018407:Bacterial Physiological Phenomena; D001436:Bacteriorhodopsins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D002712:Chlorides / Q000378:metabolism; D005051:Eubacterium / Q000378:metabolism* / Q000528:radiation effects; D025602:Halorhodopsins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D016623:Ion Pumps / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D017136:Ion Transport / Q000528:radiation effects*; D008027:Light; D009154:Mutation / Q000235:genetics; D010802:Phylogeny; D012964:Sodium / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D002712:Chlorides; D025602:Halorhodopsins; D016623:Ion Pumps; D001436:Bacteriorhodopsins; D012964:Sodium
keywords: biophysics; chloride transport; mutant; photoreceptor; proton pump; rhodopsin; sodium transport