Rhodopsin-bestrophin fusion proteins from unicellular algae form gigantic pentameric ion channels

Andrey Rozenberg, Igor Kaczmarczyk, Donna Matzov, Johannes Vierock, Takashi Nagata, Masahiro Sugiura, Kota Katayama, Yuma Kawasaki, Masae Konno, Yujiro Nagasaka, Mako Aoyama, Ishita Das, Efrat Pahima, Jonathan Church, Suliman Adam, Veniamin A. Borin, Ariel Chazan, Sandra Augustin, Jonas Wietek, Julien DineYoav Peleg, Akira Kawanabe, Yuichiro Fujiwara, Ofer Yizhar, Mordechai Sheves, Igor Schapiro, Yuji Furutani, Hideki Kandori, Keiichi Inoue, Peter Hegemann, Oded Béjà, Moran Shalev-Benami

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Many organisms sense light using rhodopsins, photoreceptive proteins containing a retinal chromophore. Here we report the discovery, structure and biophysical characterization of bestrhodopsins, a microbial rhodopsin subfamily from marine unicellular algae, in which one rhodopsin domain of eight transmembrane helices or, more often, two such domains in tandem, are C-terminally fused to a bestrophin channel. Cryo-EM analysis of a rhodopsin-rhodopsin-bestrophin fusion revealed that it forms a pentameric megacomplex (~700 kDa) with five rhodopsin pseudodimers surrounding the channel in the center. Bestrhodopsins are metastable and undergo photoconversion between red- and green-absorbing or green- and UVA-absorbing forms in the different variants. The retinal chromophore, in a unique binding pocket, photoisomerizes from all-trans to 11-cis form. Heterologously expressed bestrhodopsin behaves as a light-modulated anion channel.

Original languageEnglish
Pages (from-to)592-603
Number of pages12
JournalNature Structural and Molecular Biology
Volume29
Issue number6
DOIs
StatePublished - Jun 2022
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.

Funding

We thank all the research initiatives that produced sequencing data used in this study, D. Bleiberg and S. Larom from the Faculty of Biology at the Technion, N. Elad at the electron microscopy unit at the Weizmann Institute of Science, and the staff at the Research Instrument and Equipment Center, Kagawa University, for their technical support. This work was supported by the Israel Science Foundation (F.I.R.S.T. program no. 3592/19 to O. B. and a Research Center grant no. 3131/20 to O. B., I. S., and O. Y.), the Kimmelman center for Biomolecular Structure and Assembly (M. Sheves), Grants-in-Aid from the Japan Society for the Promotion of Science (JSPS) for Scientific Research (KAKENHI grant nos. 17H03007 and 20K21383 to K. I.; 20K21416 to T. N.; 18H03986 and 21H04969 to H. K.), Grant-in-Aid for Transformative Research areas (b) ‘Low-Energy Manipulation’ from MEXT, Japan (KAKENHI grant no. 20H05758 to K. I.), Grant-in-Aid for Scientific Research on Innovative Areas ‘Non-equilibrium-state molecular movies and their applications (Molecular Movies)’ from MEXT, Japan (KAKENHI grant no. 19H05784 to Y. Furutani) and the Japan Science and Technology Agency (JST), Japan, PRESTO (grant nos. JPMJPR1888 to T. N.; JPMJPR1903 to M. K.; and JPMJPR19G4 to K. K.), CREST (grant nos. JPMJCR1753 to H. K; and JPMJCR17N5 to Y. Furuani), Takeda Science Foundation (Y. Fujiwara), German Research Foundation (SPP1926 no. 425994138 to P. H.), the Zuckerman STEM Leadership Program (M. S.-B.), the Yeda-Sela-SABRA-WRC grant (M. S.-B.), the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (No. 949364 to M. S.-B., and 693742 to P. H.) and the German research foundation DFG (SFB1315 to P. H.). P. H. is Hertie Professor for Biophysics and is supported by the Hertie Foundation, J. D. is incumbent of the Achar Research Fellow Chair in Electrophysiology, M. Sheves holds the Katzir-Makineni Chair in Chemistry, O. B. holds the Louis and Lyra Richmond Chair in Life Sciences, and M. S.-B. holds the Tauro Career Development Chair in Biomedical Research. Y. Furuani is supported by the Equipment Sharing Division in the Organization for Co-Creation Research and Social Contributions in Nagoya Institute of Technology.

FundersFunder number
Hertie Foundation
Kimmelman center for Biomolecular Structure and Assembly
Takeda Science Foundation
Horizon 2020 Framework Programme693742, 949364
Kagawa University
European Commission
Deutsche Forschungsgemeinschaft425994138, SFB1315, SPP1926
Japan Society for the Promotion of Science20K21416, 20K21383, 21H04969, 17H03007, 18H03986
Ministry of Education, Culture, Sports, Science and Technology19H05784, 20H05758
Weizmann Institute of Science
Japan Science and Technology Agency
Core Research for Evolutional Science and TechnologyJPMJCR1753, JPMJCR17N5
Israel Science Foundation3131/20, 3592/19
Technion-Israel Institute of Technology
Precursory Research for Embryonic Science and TechnologyJPMJPR1903, JPMJPR19G4, JPMJPR1888

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