Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization

Abstract Microbial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorgani...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Masuzu Kikuchi, Keiichi Kojima, Shin Nakao, Susumu Yoshizawa, Shiho Kawanishi, Atsushi Shibukawa, Takashi Kikukawa, Yuki Sudo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/364fd69ca7db4c9e8dd68a338b8bba71
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:364fd69ca7db4c9e8dd68a338b8bba71
record_format	dspace
spelling	oai:doaj.org-article:364fd69ca7db4c9e8dd68a338b8bba712021-12-02T16:50:24ZFunctional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization10.1038/s41598-021-94181-w2045-2322https://doaj.org/article/364fd69ca7db4c9e8dd68a338b8bba712021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94181-whttps://doaj.org/toc/2045-2322Abstract Microbial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pK a = 3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins.Masuzu KikuchiKeiichi KojimaShin NakaoSusumu YoshizawaShiho KawanishiAtsushi ShibukawaTakashi KikukawaYuki SudoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Masuzu Kikuchi Keiichi Kojima Shin Nakao Susumu Yoshizawa Shiho Kawanishi Atsushi Shibukawa Takashi Kikukawa Yuki Sudo Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
description	Abstract Microbial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pK a = 3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins.
format	article
author	Masuzu Kikuchi Keiichi Kojima Shin Nakao Susumu Yoshizawa Shiho Kawanishi Atsushi Shibukawa Takashi Kikukawa Yuki Sudo
author_facet	Masuzu Kikuchi Keiichi Kojima Shin Nakao Susumu Yoshizawa Shiho Kawanishi Atsushi Shibukawa Takashi Kikukawa Yuki Sudo
author_sort	Masuzu Kikuchi
title	Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
title_short	Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
title_full	Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
title_fullStr	Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
title_full_unstemmed	Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization
title_sort	functional expression of the eukaryotic proton pump rhodopsin omr2 in escherichia coli and its photochemical characterization
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/364fd69ca7db4c9e8dd68a338b8bba71
work_keys_str_mv	AT masuzukikuchi functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT keiichikojima functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT shinnakao functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT susumuyoshizawa functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT shihokawanishi functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT atsushishibukawa functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT takashikikukawa functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization AT yukisudo functionalexpressionoftheeukaryoticprotonpumprhodopsinomr2inescherichiacolianditsphotochemicalcharacterization
_version_	1718383030279602176

Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization

Ejemplares similares