WFRF:(Engqvist R)
Sökning: WFRF:(Engqvist R) > Directed evolution ...
Directed evolution of gloeobacter violaceus rhodopsin spectral properties
- Engqvist, Martin, 1983 (författare)
- McIsaac, R.S. (författare)
- Dollinger, P. (författare)
- visa fler...
- Flytzanis, N.C. (författare)
- Abrams, M. (författare)
- Schor, S. (författare)
- Arnold, F.H. (författare)
- visa färre...
- Elsevier BV, 2015
- 2015
- Engelska.
- Ingår i: Journal of Molecular Biology. - : Elsevier BV. - 0022-2836 .- 1089-8638. ; 427:1, s. 205-220
- Tidskriftsartikel (refereegranskat)
Abstract
Ämnesord
Stäng
- Proton-pumping rhodopsins (PPRs) are photoactive retinal-binding proteins that transport ions across biological membranes in response to light. These proteins are interesting for light-harvesting applications in bioenergy production, in optogenetics applications in neuroscience, and as fluorescent sensors of membrane potential. Little is known, however, about how the protein sequence determines the considerable variation in spectral properties of PPRs from different biological niches or how to engineer these properties in a given PPR. Here we report a comprehensive study of amino acid substitutions in the retinal-binding pocket of Gloeobacter violaceus rhodopsin (GR) that tune its spectral properties. Directed evolution generated 70 GR variants with absorption maxima shifted by up to ± 80 nm, extending the protein's light absorption significantly beyond the range of known natural PPRs. While proton-pumping activity was disrupted in many of the spectrally shifted variants, we identified single tuning mutations that incurred blue and red shifts of 42 nm and 22 nm, respectively, that did not disrupt proton pumping. Blue-shifting mutations were distributed evenly along the retinal molecule while red-shifting mutations were clustered near the residue K257, which forms a covalent bond with retinal through a Schiff base linkage. Thirty eight of the identified tuning mutations are not found in known microbial rhodopsins. We discovered a subset of red-shifted GRs that exhibit high levels of fluorescence relative to the WT (wild-type) protein.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- opsins
- optogenetics
- proton pumps
- fluorescent proteins
- Retinal
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
Hitta via bibliotek
- Journal of Molecular Biology (Sök värdpublikationen i LIBRIS)
Till lärosätets databas
Hitta mer i SwePub
- Av författaren/redakt...
- Engqvist, Martin ...
- McIsaac, R.S.
- Dollinger, P.
- Flytzanis, N.C.
- Abrams, M.
- Schor, S.
- visa fler...
- Arnold, F.H.
- visa färre...
- Om ämnet
-
- NATURVETENSKAP
- NATURVETENSKAP
- och Biologi
- och Biokemi och mole ...
- Artiklar i publikationen
- Journal of Molec ...
- Av lärosätet
- Chalmers tekniska högskola
Sök utanför SwePub
- Sök vidare i:
- Google Book Search
- Google Scholar
Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.
- Copyright © LIBRIS - Nationella bibliotekssystem
- LIBRIS.kb.se
