Selection and Characterization of Artificial Proteins Targeting the Tubulin α Subunit.
artificial protein
in vitro selection
microtubule
tubulin
αRep
Journal
Structure (London, England : 1993)
ISSN: 1878-4186
Titre abrégé: Structure
Pays: United States
ID NLM: 101087697
Informations de publication
Date de publication:
05 03 2019
05 03 2019
Historique:
received:
26
07
2018
revised:
24
10
2018
accepted:
03
12
2018
pubmed:
22
1
2019
medline:
13
3
2020
entrez:
22
1
2019
Statut:
ppublish
Résumé
Microtubules are cytoskeletal filaments of eukaryotic cells made of αβ-tubulin heterodimers. Structural studies of non-microtubular tubulin rely mainly on molecules that prevent its self-assembly and are used as crystallization chaperones. Here we identified artificial proteins from an αRep library that are specific to α-tubulin. Turbidity experiments indicate that these αReps impede microtubule assembly in a dose-dependent manner and total internal reflection fluorescence microscopy further shows that they specifically block growth at the microtubule (-) end. Structural data indicate that they do so by targeting the α-tubulin longitudinal surface. Interestingly, in one of the complexes studied, the α subunit is in a conformation that is intermediate between the ones most commonly observed in X-ray structures of tubulin and those seen in the microtubule, emphasizing the plasticity of tubulin. These α-tubulin-specific αReps broaden the range of tools available for the mechanistic study of microtubule dynamics and its regulation.
Identifiants
pubmed: 30661854
pii: S0969-2126(18)30457-X
doi: 10.1016/j.str.2018.12.001
pmc: PMC6408325
pii:
doi:
Substances chimiques
Peptide Library
0
Recombinant Fusion Proteins
0
Tubulin
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
497-506.e4Subventions
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Medical Research Council
ID : FC001163
Pays : United Kingdom
Organisme : Cancer Research UK
ID : FC001163
Pays : United Kingdom
Organisme : Wellcome Trust
ID : FC001163
Pays : United Kingdom
Informations de copyright
Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.
Références
Nat Cell Biol. 2015 Nov;17(11):1422-34
pubmed: 26414402
J Struct Biol. 2018 Feb;201(2):88-99
pubmed: 28823563
Science. 2012 Aug 17;337(6096):857-60
pubmed: 22904013
Nat Cell Biol. 2014 Aug;16(8):804-11
pubmed: 24997520
J Mol Biol. 2011 Sep 9;412(1):35-42
pubmed: 21787788
Protein Sci. 2011 Mar;20(3):482-91
pubmed: 21213247
J Cell Biol. 2017 Sep 4;216(9):2669-2677
pubmed: 28652389
Biochemistry. 2005 Nov 8;44(44):14616-25
pubmed: 16262261
Annu Rev Cell Dev Biol. 1997;13:83-117
pubmed: 9442869
Cell. 2000 Sep 15;102(6):809-16
pubmed: 11030624
Nat Commun. 2018 Apr 25;9(1):1662
pubmed: 29695795
Methods Enzymol. 1991;196:478-85
pubmed: 2034137
Dev Cell. 2016 May 23;37(4):362-376
pubmed: 27219064
J Biol Chem. 2014 Sep 5;289(36):25199-210
pubmed: 25056950
PLoS One. 2013 Aug 27;8(8):e71512
pubmed: 24014183
Methods Mol Med. 2007;137:235-43
pubmed: 18085233
J Biol Chem. 2012 Apr 27;287(18):15143-53
pubmed: 22403406
J Am Chem Soc. 2014 Sep 10;136(36):12615-23
pubmed: 25162583
Cell Struct Funct. 1999 Oct;24(5):299-303
pubmed: 15216886
Nat Struct Mol Biol. 2018 Jul;25(7):607-615
pubmed: 29967541
Nat Commun. 2014 Nov 14;5:5364
pubmed: 25395082
Proc Natl Acad Sci U S A. 2018 Jul 3;115(27):E6191-E6200
pubmed: 29915050
Protein Expr Purif. 2003 Nov;32(1):83-8
pubmed: 14680943
Elife. 2014 Aug 05;3:e03069
pubmed: 25097237
Biochemistry. 1997 Sep 9;36(36):10817-21
pubmed: 9312271
J Biol Chem. 2012 Sep 7;287(37):31085-94
pubmed: 22791712
Methods Cell Biol. 2010;95:555-80
pubmed: 20466153
Nat Struct Mol Biol. 2017 Nov;24(11):931-943
pubmed: 28991265
J Mol Biol. 2001 Nov 9;313(5):1045-57
pubmed: 11700061
J Am Chem Soc. 2015 Jul 29;137(29):9218-21
pubmed: 26165802
FEBS Lett. 2013 Nov 1;587(21):3450-5
pubmed: 24021646
Sci Rep. 2016 Jul 06;6:28922
pubmed: 27380724
Curr Biol. 2014 Feb 17;24(4):372-84
pubmed: 24508171
Cell. 1999 Jan 8;96(1):79-88
pubmed: 9989499
J Cell Biol. 2013 Feb 4;200(3):259-70
pubmed: 23358242
Nat Cell Biol. 2016 Oct;18(10):1102-8
pubmed: 27617931
Annu Rev Pharmacol Toxicol. 2015;55:489-511
pubmed: 25562645
J Biol Chem. 2016 Jun 17;291(25):12907-15
pubmed: 27129203
Biochemistry. 1989 Nov 14;28(23):9143-52
pubmed: 2605248
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
pubmed: 20383002
Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10037-41
pubmed: 11517324
Biochemistry. 2011 Oct 11;50(40):8636-44
pubmed: 21888381
Nat Struct Mol Biol. 2013 Aug;20(8):1001-7
pubmed: 23872990
Cell. 2015 Aug 13;162(4):849-59
pubmed: 26234155
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
pubmed: 20124692
Nature. 2004 Mar 11;428(6979):198-202
pubmed: 15014504
Science. 2013 Feb 1;339(6119):587-90
pubmed: 23287720
J Mol Biol. 2010 Nov 26;404(2):307-27
pubmed: 20887736
Proc Natl Acad Sci U S A. 2012 Jul 24;109(30):12011-6
pubmed: 22778434
Dev Cell. 2016 Apr 4;37(1):72-84
pubmed: 27046833
Nat Commun. 2017 Jul 10;8(1):70
pubmed: 28694425
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
pubmed: 19461840
Curr Opin Struct Biol. 2017 Oct;46:65-70
pubmed: 28628789