Single-molecule FRET and conformational analysis of beta-arrestin-1 through genetic code expansion and a Se-click reaction.
Journal
Chemical science
ISSN: 2041-6520
Titre abrégé: Chem Sci
Pays: England
ID NLM: 101545951
Informations de publication
Date de publication:
07 Jul 2021
07 Jul 2021
Historique:
received:
14
05
2021
revised:
02
07
2021
accepted:
27
05
2021
entrez:
19
7
2021
pubmed:
20
7
2021
medline:
20
7
2021
Statut:
epublish
Résumé
Single-molecule Förster resonance energy transfer (smFRET) is a powerful tool for investigating the dynamic properties of biomacromolecules. However, the success of protein smFRET relies on the precise and efficient labeling of two or more fluorophores on the protein of interest (POI), which has remained highly challenging, particularly for large membrane protein complexes. Here, we demonstrate the site-selective incorporation of a novel unnatural amino acid (2-amino-3-(4-hydroselenophenyl) propanoic acid, SeF) through genetic expansion followed by a Se-click reaction to conjugate the Bodipy593 fluorophore on calmodulin (CaM) and β-arrestin-1 (βarr1). Using this strategy, we monitored the subtle but functionally important conformational change of βarr1 upon activation by the G-protein coupled receptor (GPCR) through smFRET for the first time. Our new method has broad applications for the site-specific labeling and smFRET measurement of membrane protein complexes, and the elucidation of their dynamic properties such as transducer protein selection.
Identifiants
pubmed: 34276941
doi: 10.1039/d1sc02653d
pii: d1sc02653d
pmc: PMC8261736
doi:
Types de publication
Journal Article
Langues
eng
Pagination
9114-9123Commentaires et corrections
Type : ErratumIn
Informations de copyright
This journal is © The Royal Society of Chemistry.
Déclaration de conflit d'intérêts
There are no conflicts to declare.
Références
Angew Chem Int Ed Engl. 2012 Apr 23;51(17):4166-70
pubmed: 22473599
Mol Cell. 2011 May 6;42(3):367-77
pubmed: 21549313
Nature. 2015 Oct 29;526(7575):687-91
pubmed: 26511579
Biopolymers. 2008 Jul;89(7):565-77
pubmed: 18286627
PLoS One. 2014 Feb 04;9(2):e87649
pubmed: 24503716
Angew Chem Int Ed Engl. 2002 Jul 15;41(14):2596-9
pubmed: 12203546
J Am Chem Soc. 2013 Jan 30;135(4):1558-69
pubmed: 23272641
Nature. 2019 Apr;568(7752):415-419
pubmed: 30971821
Science. 2020 Jan 3;367(6473):76-79
pubmed: 31896714
Proc Natl Acad Sci U S A. 2018 Apr 10;115(15):3834-3839
pubmed: 29581292
Nat Methods. 2019 Sep;16(9):815-829
pubmed: 31471616
Angew Chem Int Ed Engl. 2009;48(38):6974-98
pubmed: 19714693
Nat Chem Biol. 2007 Jun;3(6):321-2
pubmed: 17450134
Nature. 2017 Jul 6;547(7661):68-73
pubmed: 28607487
Cell. 2019 Jan 24;176(3):468-478.e11
pubmed: 30639099
Science. 2018 Jan 19;359(6373):
pubmed: 29348210
J Phys Chem B. 2016 May 19;120(19):4357-64
pubmed: 27111039
Cell. 2015 Oct 22;163(3):734-45
pubmed: 26456112
Nat Chem. 2016 Feb;8(2):120-8
pubmed: 26791894
Science. 2017 Feb 10;355(6325):597-602
pubmed: 28183972
J Med Chem. 2004 Mar 25;47(7):1739-49
pubmed: 15027865
Nat Methods. 2005 Mar;2(3):171-6
pubmed: 15782185
Biochem Soc Trans. 2017 Jun 15;45(3):759-769
pubmed: 28620037
Nat Methods. 2018 Nov;15(11):984
pubmed: 30327572
Nature. 2019 May;569(7757):514-518
pubmed: 31092918
Nature. 2013 May 2;497(7447):137-41
pubmed: 23604254
Chem Rev. 2017 Jan 11;117(1):186-245
pubmed: 27341004
Angew Chem Int Ed Engl. 2017 Dec 11;56(50):15818-15827
pubmed: 28857389
Science. 2000 Jun 16;288(5473):2048-51
pubmed: 10856219
Nat Commun. 2017 Feb 09;8:14335
pubmed: 28181498
Nat Commun. 2015 Sep 08;6:8202
pubmed: 26347956
Chem Rev. 2018 Jan 10;118(1):119-141
pubmed: 28627171
Acc Chem Res. 2011 Sep 20;44(9):828-39
pubmed: 21609129
Biochemistry. 2006 Dec 5;45(48):14233-46
pubmed: 17128963
Angew Chem Int Ed Engl. 2020 Jul 27;59(31):12885-12893
pubmed: 32342666
J Am Chem Soc. 2019 May 15;141(19):7698-7703
pubmed: 31038942
Elife. 2018 Feb 02;7:
pubmed: 29393851
Nat Rev Microbiol. 2013 May;11(5):303-15
pubmed: 23549067
Nature. 2017 Oct 26;550(7677):543-547
pubmed: 29045395
Nature. 2020 Mar;579(7798):303-308
pubmed: 31945771
Science. 1991 Oct 4;254(5028):51-8
pubmed: 1925561
Science. 2020 Feb 21;367(6480):881-887
pubmed: 32079767
Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6264-8
pubmed: 8692803
Nature. 2020 Mar;579(7798):297-302
pubmed: 31945772
Science. 2003 Jun 27;300(5628):2061-5
pubmed: 12791999
J Am Chem Soc. 2011 Mar 2;133(8):2463-80
pubmed: 21291253
Biochim Biophys Acta Biomembr. 2017 Jul;1859(7):1242-1253
pubmed: 28392350
J Am Chem Soc. 2012 Jun 27;134(25):10317-20
pubmed: 22694658
Nat Protoc. 2008;3(9):1527-34
pubmed: 18772880
Science. 2019 Jul 5;365(6448):61-65
pubmed: 31273117
Proc Natl Acad Sci U S A. 2015 Nov 17;112(46):14254-9
pubmed: 26578769
Structure. 2019 Jul 2;27(7):1162-1170.e3
pubmed: 31080119
Annu Rev Biochem. 2010;79:413-44
pubmed: 20307192
Cell. 2020 Dec 23;183(7):1813-1825.e18
pubmed: 33296703
J Am Chem Soc. 2020 Oct 7;142(40):17057-17068
pubmed: 32915556
Nat Chem Biol. 2018 Sep;14(9):876-886
pubmed: 30120361
Mol Biosyst. 2010 Sep;6(9):1540-7
pubmed: 20601974