Structural basis for channel conduction in the pump-like channelrhodopsin ChRmine.
ChRmine
PLCR
all-optical
channelrhodopsin
cryo-EM
microbial opsin
optogenetics
pump-like channelrhodopsin
structure-guided engineering
Journal
Cell
ISSN: 1097-4172
Titre abrégé: Cell
Pays: United States
ID NLM: 0413066
Informations de publication
Date de publication:
17 02 2022
17 02 2022
Historique:
received:
31
10
2021
revised:
13
12
2021
accepted:
11
01
2022
pubmed:
4
2
2022
medline:
11
3
2022
entrez:
3
2
2022
Statut:
ppublish
Résumé
ChRmine, a recently discovered pump-like cation-conducting channelrhodopsin, exhibits puzzling properties (large photocurrents, red-shifted spectrum, and extreme light sensitivity) that have created new opportunities in optogenetics. ChRmine and its homologs function as ion channels but, by primary sequence, more closely resemble ion pump rhodopsins; mechanisms for passive channel conduction in this family have remained mysterious. Here, we present the 2.0 Å resolution cryo-EM structure of ChRmine, revealing architectural features atypical for channelrhodopsins: trimeric assembly, a short transmembrane-helix 3, a twisting extracellular-loop 1, large vestibules within the monomer, and an opening at the trimer interface. We applied this structure to design three proteins (rsChRmine and hsChRmine, conferring further red-shifted and high-speed properties, respectively, and frChRmine, combining faster and more red-shifted performance) suitable for fundamental neuroscience opportunities. These results illuminate the conduction and gating of pump-like channelrhodopsins and point the way toward further structure-guided creation of channelrhodopsins for applications across biology.
Identifiants
pubmed: 35114111
pii: S0092-8674(22)00031-9
doi: 10.1016/j.cell.2022.01.007
pmc: PMC7612760
mid: EMS145033
pii:
doi:
Substances chimiques
Channelrhodopsins
0
Schiff Bases
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
672-689.e23Subventions
Organisme : NIMH NIH HHS
ID : R01 MH075957
Pays : United States
Organisme : Medical Research Council
ID : MC_UP_A025_1012
Pays : United Kingdom
Commentaires et corrections
Type : CommentIn
Informations de copyright
Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The ChRmine antibody (Fab02) and ChRmine variants are described in the pending patent application material; these tools and all methods, protocols, clones, and sequences are freely available to nonprofit institutions and investigators. K.D. is a member of the Cell advisory board.
Références
J Struct Biol. 2015 Nov;192(2):216-21
pubmed: 26278980
Nat Methods. 2014 Mar;11(3):338-46
pubmed: 24509633
Sci Rep. 2015 Nov 24;5:16450
pubmed: 26597707
Mol Biol Evol. 2016 Jul;33(7):1870-4
pubmed: 27004904
Proc Natl Acad Sci U S A. 2016 Jan 26;113(4):822-9
pubmed: 26699459
Nat Commun. 2018 May 1;9(1):1750
pubmed: 29717130
Methods Mol Biol. 2018;1700:97-109
pubmed: 29177828
Nat Commun. 2013;4:1678
pubmed: 23575682
Biophys Physicobiol. 2017 May 20;14:57-66
pubmed: 28630812
Acta Crystallogr D Struct Biol. 2021 Oct 1;77(Pt 10):1282-1291
pubmed: 34605431
mBio. 2020 Apr 21;11(2):
pubmed: 32317325
Cell. 2019 May 16;177(5):1346-1360.e24
pubmed: 31080068
PLoS One. 2014 Jun 05;9(6):e98924
pubmed: 24901492
Science. 2017 Sep 15;357(6356):
pubmed: 28912215
Nature. 2021 Aug;596(7873):590-596
pubmed: 34293799
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
pubmed: 21460454
Science. 2014 Apr 25;344(6182):420-4
pubmed: 24763591
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):13940-5
pubmed: 14615590
Neuron. 2020 Sep 9;107(5):836-853.e11
pubmed: 32574559
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
J Neural Eng. 2021 Jul 26;18(4):
pubmed: 34229315
J Mol Biol. 2010 Feb 26;396(3):564-79
pubmed: 19961859
Science. 1997 Sep 12;277(5332):1676-81
pubmed: 9287223
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
pubmed: 20057044
Sci Rep. 2018 Sep 3;8(1):13123
pubmed: 30177765
IUCrJ. 2020 Feb 11;7(Pt 2):253-267
pubmed: 32148853
Nat Med. 2021 Jul;27(7):1223-1229
pubmed: 34031601
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W665-7
pubmed: 15215472
Acta Crystallogr D Biol Crystallogr. 2013 Oct;69(Pt 10):1965-80
pubmed: 24100316
Science. 2019 Aug 9;365(6453):
pubmed: 31320556
Biophys Rev. 2020 Apr;12(2):355-361
pubmed: 32065378
Neuron. 2022 Apr 6;110(7):1139-1155.e6
pubmed: 35120626
Science. 2015 Aug 7;349(6248):590-1
pubmed: 26250674
Nature. 2019 Oct;574(7776):132-136
pubmed: 31554965
Science. 2000 May 26;288(5470):1390-6
pubmed: 10827943
Proteins. 2013 Sep;81(9):1585-92
pubmed: 23625688
Nat Neurosci. 2015 Sep;18(9):1213-25
pubmed: 26308982
IUCrJ. 2019 Jan 01;6(Pt 1):5-17
pubmed: 30713699
Structure. 2012 Apr 4;20(4):582-92
pubmed: 22483106
Mol Biol Evol. 1987 Jul;4(4):406-25
pubmed: 3447015
Biochemistry. 1988 Nov 15;27(23):8516-20
pubmed: 2851326
J Mol Biol. 2006 May 5;358(3):675-85
pubmed: 16540121
J Am Chem Soc. 2015 Mar 11;137(9):3291-9
pubmed: 25712566
Nature. 2021 Aug;596(7873):583-589
pubmed: 34265844
Sci Signal. 2019 Mar 19;12(573):
pubmed: 30890657
Nature. 2015 May 7;521(7550):48-53
pubmed: 25849775
Biophys Physicobiol. 2015 Dec 11;12:121-9
pubmed: 27493861
Nature. 2002 Oct 3;419(6906):484-7
pubmed: 12368857
Elife. 2021 Mar 23;10:
pubmed: 33752801
Cell. 2011 Dec 23;147(7):1446-57
pubmed: 22196724
J Biol Chem. 2012 Sep 14;287(38):31804-12
pubmed: 22843694
Nat Commun. 2015 May 15;6:7177
pubmed: 25975962
Biophys J. 2016 Jun 7;110(11):2302-2304
pubmed: 27233115
Proc Natl Acad Sci U S A. 2017 Nov 7;114(45):E9512-E9519
pubmed: 29078348
Nat Methods. 2016 Apr;13(4):325-8
pubmed: 26878381
Chem Rev. 2014 Jan 8;114(1):126-63
pubmed: 24364740
Science. 2017 Nov 24;358(6366):
pubmed: 29170206
Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10037-41
pubmed: 11517324
Biophys J. 2015 Jun 2;108(11):2680-90
pubmed: 26039169
Nat Struct Mol Biol. 2016 Jun;23(6):481-6
pubmed: 27273631
J Struct Biol. 2005 Oct;152(1):36-51
pubmed: 16182563
Nat Neurosci. 2008 Jun;11(6):631-3
pubmed: 18432196
Adv Exp Med Biol. 2021;1293:35-53
pubmed: 33398806
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Proc Natl Acad Sci U S A. 2013 Apr 2;110(14):E1273-81
pubmed: 23509282
Nat Commun. 2017 Nov 10;8(1):1430
pubmed: 29127285
Nature. 2018 Sep;561(7723):349-354
pubmed: 30158697
Nature. 2000 Aug 10;406(6796):569-70
pubmed: 10949279
Proc Natl Acad Sci U S A. 2021 Apr 6;118(14):
pubmed: 33790007
Nat Commun. 2018 Sep 26;9(1):3949
pubmed: 30258177
Nat Commun. 2019 Jul 25;10(1):3315
pubmed: 31346176
Elife. 2018 Nov 09;7:
pubmed: 30412051
Structure. 2006 Apr;14(4):673-81
pubmed: 16615909
Science. 2016 Dec 23;354(6319):1552-1557
pubmed: 28008064
Bioinformatics. 2013 Nov 1;29(21):2722-8
pubmed: 23986568
PLoS One. 2014 Sep 30;9(9):e108362
pubmed: 25268964
J Struct Biol. 2020 Aug 1;211(2):107545
pubmed: 32534144
Sci Rep. 2019 Aug 2;9(1):11283
pubmed: 31375689
Proc Natl Acad Sci U S A. 1990 Dec 15;87(24):9774-8
pubmed: 11607137
J Mol Biol. 2002 May 31;319(2):555-65
pubmed: 12051928
Cell. 2022 Feb 17;185(4):672-689.e23
pubmed: 35114111
Nat Commun. 2020 May 1;11(1):2137
pubmed: 32358514
Science. 2019 Jul 5;365(6448):61-65
pubmed: 31273117
PLoS One. 2015 Jun 26;10(6):e0131094
pubmed: 26114863
Nature. 2012 Jan 22;482(7385):369-74
pubmed: 22266941
Nat Biotechnol. 2021 Feb;39(2):161-164
pubmed: 33020604
Nature. 2015 Oct 29;526(7575):653-9
pubmed: 26436451
J Mol Biol. 2008 Feb 1;375(5):1267-81
pubmed: 18082767
Curr Biol. 2020 Dec 21;30(24):4910-4920.e5
pubmed: 33065010
Nat Methods. 2017 Apr;14(4):331-332
pubmed: 28250466
EMBO J. 2000 May 15;19(10):2152-60
pubmed: 10811606
Cell. 2021 Oct 14;184(21):5279-5285
pubmed: 34562367
Nature. 2018 Sep;561(7723):343-348
pubmed: 30158696
Science. 2002 Jun 28;296(5577):2395-8
pubmed: 12089443
Protein Sci. 2018 Jan;27(1):14-25
pubmed: 28710774
Nat Neurosci. 2010 Mar;13(3):387-92
pubmed: 20081849
Nucleic Acids Res. 2014 Jul;42(Web Server issue):W320-4
pubmed: 24753421
Sci Rep. 2018 May 29;8(1):8262
pubmed: 29844455
Nucleic Acids Res. 2008 Apr;36(7):2295-300
pubmed: 18287115
Science. 2014 Apr 25;344(6182):409-12
pubmed: 24674867
Proc Natl Acad Sci U S A. 2021 Mar 30;118(13):
pubmed: 33753488
Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16561-5
pubmed: 18922772