Light-coupled cryo-plunger for time-resolved cryo-EM.
Cryo-EM
Cryo-electron microscopy
Manual cryo-plunger
Single particle
Time-resolved
Journal
Journal of structural biology
ISSN: 1095-8657
Titre abrégé: J Struct Biol
Pays: United States
ID NLM: 9011206
Informations de publication
Date de publication:
01 12 2020
01 12 2020
Historique:
received:
05
06
2020
revised:
08
09
2020
accepted:
11
09
2020
pubmed:
21
9
2020
medline:
15
10
2021
entrez:
20
9
2020
Statut:
ppublish
Résumé
Proteins are dynamic molecules that can undergo rapid conformational rearrangements in response to stimuli. These structural changes are often critical to protein function, and thus elucidating time-dependent conformational landscapes has been a long-standing goal of structural biology. To harness the power of single particle cryo-EM methods to enable 'time-resolved' structure determination, we have developed a light-coupled cryo-plunger that pairs flash-photolysis of caged ligands with rapid sample vitrification. The 'flash-plunger' consists of a high-power ultraviolet LED coupled with focusing optics and a motorized linear actuator, enabling the user to immobilize protein targets in vitreous ice within a programmable time window - as short as tens of milliseconds - after stimulus delivery. The flash-plunger is a simple, inexpensive and flexible tool to explore short-lived conformational states previously unobtainable by conventional sample preparation methods.
Identifiants
pubmed: 32950604
pii: S1047-8477(20)30197-0
doi: 10.1016/j.jsb.2020.107624
pmc: PMC7959588
mid: NIHMS1636071
pii:
doi:
Substances chimiques
Ligands
0
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
107624Subventions
Organisme : NINDS NIH HHS
ID : R01 NS038631
Pays : United States
Organisme : NIH HHS
ID : DP5 OD017871
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : NIDDK NIH HHS
ID : T32 DK007680
Pays : United States
Organisme : NIMH NIH HHS
ID : F32 MH115595
Pays : United States
Organisme : NIGMS NIH HHS
ID : U24 GM129539
Pays : United States
Organisme : NIGMS NIH HHS
ID : U24 GM129547
Pays : United States
Organisme : NINDS NIH HHS
ID : F31 NS096782
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.
Références
Ultramicroscopy. 2019 Aug;203:170-180
pubmed: 30528101
Elife. 2018 Aug 30;7:
pubmed: 30160233
ACS Chem Neurosci. 2013 Aug 21;4(8):1163-7
pubmed: 23672485
J Neurosci Methods. 2004 Feb 15;133(1-2):153-9
pubmed: 14757356
Physiol Rev. 2002 Jul;82(3):735-67
pubmed: 12087134
Front Mol Biosci. 2016 Sep 09;3:47
pubmed: 27668215
Ultramicroscopy. 1994 Dec;56(4):241-52
pubmed: 7831735
BMC Biol. 2018 May 31;16(1):55
pubmed: 29848334
Nature. 2013 Dec 5;504(7478):113-8
pubmed: 24305161
Science. 2014 Aug 29;345(6200):1021-6
pubmed: 25103405
J Neurosci. 2011 Nov 16;31(46):16550-5
pubmed: 22090481
Anal Chem. 2010 Sep 1;82(17):7422-9
pubmed: 20712348
Nature. 2007 Sep 20;449(7160):316-23
pubmed: 17882215
J Struct Biol. 1999 Dec 1;128(1):19-25
pubmed: 10600554
Elife. 2018 Mar 07;7:
pubmed: 29513216
Structure. 2015 Jun 2;23(6):1097-105
pubmed: 26004440
Biochemistry. 1978 May 16;17(10):1929-35
pubmed: 148906
Elife. 2020 Jun 04;9:
pubmed: 32496192
Biophys J. 2007 Nov 1;93(9):3092-104
pubmed: 17615289
J Struct Biol. 2016 Jan;193(1):1-12
pubmed: 26592709
Nat Methods. 2017 Mar;14(3):290-296
pubmed: 28165473
FEBS Lett. 2003 Nov 27;555(1):91-5
pubmed: 14630325
Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8752-6
pubmed: 8090718
J Neurosci. 2007 Aug 1;27(31):8344-57
pubmed: 17670981
J Gen Physiol. 1935 Nov 20;19(2):199-210
pubmed: 19872919
Nature. 2018 Mar 15;555(7696):397-401
pubmed: 29513651
Biochemistry. 2000 May 9;39(18):5500-8
pubmed: 10820023
Nature. 2016 May 18;534(7607):347-51
pubmed: 27281200
J Struct Biol. 2009 May;166(2):205-13
pubmed: 19374019
Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18799-804
pubmed: 17132736
Biochemistry. 1992 Jun 23;31(24):5507-14
pubmed: 1610795
J Physiol. 1910 Jul 1;40(5):404-30
pubmed: 16993016
Rev Neurosci. 2004;15(3):167-83
pubmed: 15357140
Cell. 2019 Feb 7;176(4):702-715.e14
pubmed: 30661758
Nature. 2014 Nov 13;515(7526):228-33
pubmed: 25296249
Microsc Microanal. 2008 Oct;14(5):375-9
pubmed: 18793481
Cell. 2014 Feb 13;156(4):717-29
pubmed: 24507937
Nature. 2019 Jun;570(7761):400-404
pubmed: 31108498
Nat Commun. 2019 Jun 12;10(1):2579
pubmed: 31189921
Nat Commun. 2012 Jul 03;3:936
pubmed: 22760635
Nature. 2012 Sep 20;489(7416):400-5
pubmed: 22842900
J Struct Biol. 2009 Mar;165(3):184-9
pubmed: 19114106
Elife. 2018 Jun 01;7:
pubmed: 29856314
J Electron Microsc Tech. 1988 Sep;10(1):87-111
pubmed: 3193246
Nature. 1995 Jan 5;373(6509):37-43
pubmed: 7800037
EMBO J. 1993 Jan;12(1):1-8
pubmed: 8428572
Curr Opin Neurobiol. 1998 Jun;8(3):418-24
pubmed: 9687356
J Am Chem Soc. 2005 Jul 13;127(27):9865-74
pubmed: 15998092
Elife. 2018 Nov 09;7:
pubmed: 30412051
J Struct Biol. 1998;121(3):306-13
pubmed: 9704502
Nat Neurosci. 2001 Nov;4(11):1086-92
pubmed: 11687814
Nature. 2010 Jul 15;466(7304):329-33
pubmed: 20631791
Biochemistry. 1994 Feb 15;33(6):1526-33
pubmed: 8312272
J Micromech Microeng. 2014 Nov 1;24(11):115001
pubmed: 25530679
Nat Struct Mol Biol. 2017 Oct;24(10):857-865
pubmed: 28846092
J Microsc. 2003 Jul;211(Pt 1):48-53
pubmed: 12839550
J Struct Biol. 2017 Dec;200(3):303-306
pubmed: 28625887
Nat Methods. 2017 Apr;14(4):331-332
pubmed: 28250466
Curr Opin Neurobiol. 2002 Oct;12(5):587-92
pubmed: 12367640
Nature. 2009 Jul 30;460(7255):599-604
pubmed: 19641589
J Struct Biol. 2003 Oct-Nov;144(1-2):246-52
pubmed: 14643227
Nat Neurosci. 2016 Sep;19(9):1218-24
pubmed: 27399842
Structure. 2017 Apr 4;25(4):663-670.e3
pubmed: 28286002
Nature. 2013 Dec 12;504(7479):242-247
pubmed: 24305055
J Gen Physiol. 2006 Feb;127(2):109-17
pubmed: 16418400
Methods Mol Biol. 2018;1764:59-71
pubmed: 29605908
Science. 2018 Oct 19;362(6412):
pubmed: 30049784