A natural light-driven inward proton pump

Inoue, Kazuhide; Itô, Shota; Kato, Yoshitaka; Nomura, Yurika; Shibata, Mikihiro; Uchihashi, Takayuki; Tsunoda, Satoshi P.

doi:10.1038/ncomms13415

Cited by 141 publications

(184 citation statements)

References 56 publications

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“…Blocking access of protons to the retinal Schiff base from the cell cytoplasm should enable preferential exchange of protons with the outside of the cell. This hypothesis is supported by a recent report that describes a natural light-driven inward proton pump showing the capacity of microbial rhodopsins to accept protons from outside the cell 22 . To block access of protons from the cytoplasmic side, we substituted the amino acid at the PD position of Voltron for a neutral residue (D92N).…”

supporting

confidence: 53%

A general approach to engineer positive-going eFRET voltage indicators

Abdelfattah

Valenti

Wong

et al. 2019

Preprint

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We engineered electrochromic fluorescence resonance energy transfer (eFRET) genetically encoded voltage indicators (GEVIs) with "positive-going" fluorescence response to membrane depolarization through rational manipulation of the native proton transport pathway in microbial rhodopsins. We transformed the state-of-the-art eFRET GEVI Voltron into Positron, with kinetics and sensitivity equivalent to Voltron but flipped fluorescence signal polarity. We further applied this general approach to GEVIs containing different voltage sensitive rhodopsin domains and various fluorescent dye and fluorescent protein reporters.Genetically encoded voltage indicators (GEVIs) allow visualization of fast action potentials and subthreshold dynamics in groups of genetically defined neurons with high spatiotemporal resolution 1,2 . Monitoring voltage signals in vivo in large populations of neurons will enable dissection of detailed mechanistic links between brain activity and animal behavior. Despite recent advances, no current GEVI has ideal properties for routine, robust in vivo imaging.

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supporting

confidence: 53%

A general approach to engineer positive-going eFRET voltage indicators

Abdelfattah

Valenti

Wong

et al. 2019

Preprint

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“…The widely spread presence and importance of pR-based phototrophy in the marine environment 17 was identified. Recently, rhodopsins that function as inward proton pumps were discovered 18,19 .…”

Section: Introductionmentioning

confidence: 99%

High Resolution Structural Insights into Heliorhodopsin Family

Kovalev

Volkov

Astashkin

et al. 2019

Preprint

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Rhodopsins are the most abundant light-harvesting proteins. A new family of rhodopsins, heliorhodopsins (HeRs), was recently discovered. In opposite to the known rhodopsins their N-termini face the cytoplasm. HeRs structure and function remain unknown. We present structures of two HeR-48C12 states at 1.5 Å showing its remarkable difference from all known rhodopsins. Its internal extracellular part is completely hydrophobic, while the cytoplasmic part comprises a cavity ('active site'), surrounded by charged amino acids and containing a cluster of water molecules, presumably being a primary proton acceptor from the Schiff base. At acidic pH a planar triangle molecule (acetate) is present in the 'active site' which demonstrated its ability to maintain such anions as carbonate or nitrate. Structure-based bioinformatic analysis identified 10 subfamilies of HeRs suggesting their diverse biological functions. The structures and available data suggest an enzymatic activity of HeR-48C12 subfamily and their possible involvement into fundamental redox biological processes.

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“…These results suggest that dinoflagellates use some unknown signaling mechanism that is compatible with mammalian protein targeting machinery. PR in dinoflagellates is highly similar to proteorhodopsin, which are globally abundant in bacteria (Béja et al 2013;Gómez-Consarnau et al 2010) and well characterized as a proton pump (Béja et al 2000;Inoue et al 2016). Since its first discovery in dinoflagellates (Lin et al 2010), eukaryotic homologs of PR have also been found in other eukaryotes microorganisms, such as diatoms, haptophytes and cryptophytes (Marchetti et al 2015), and recently found in arctic microbial eukaryotes (Vader et al 2018).…”

Section: Discussionmentioning

confidence: 99%

Heterologous expression and cell membrane localization of dinoflagellate rhodopsins in mammalian cells

Shi

Lin

2020

Preprint

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Rhodopsins are now found in all domains of life, and are classified into two large groups: type I, found in animals and type II found in microbes including Bacteria, Archaea, and Eukarya. While type I rhodopsin functions in many photodependent signaling processes including vision, type II among others contains rhodopsins that function as a light-driven proton pump to convert light into ATP as in proteobacteria (named proteorhodopsin). Proteorhodopsin homologs have been documented in dinoflagellates, but their subcellular localizations and functions are still poorly understood. Even though sequence analyses suggest that it is a membrane protein, experimental evidence that dinoflagellate rhodopsins are localized on the plasma membrane or endomembranes is still lacking. As no robust dinoflagellate gene transformation tool is available, we used HEK 293T cells to construct a mammalian expression system for two dinoflagellate rhodopsin genes. The success of expressing these genes in the system shows that this mammalian cell type is suitable for expressing dinoflagellate genes.Immunofluorescence of the expressed protein locates these dinoflagellate rhodopsins on the cell membrane. This result indicates that the protein codons and membrane targeting signal of the dinoflagellate genes are compatible with the mammalian cells, and the proteins' subcellular localization is consistent with proton pump rhodopsins.

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A natural light-driven inward proton pump

Cited by 141 publications

References 56 publications

A general approach to engineer positive-going eFRET voltage indicators

A general approach to engineer positive-going eFRET voltage indicators

High Resolution Structural Insights into Heliorhodopsin Family

Heterologous expression and cell membrane localization of dinoflagellate rhodopsins in mammalian cells

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