Quantifying the Monomer-Dimer Equilibrium of Tubulin with Mass Photometry.
binding affinity
mass photometry
single molecule
tubulin
Journal
Journal of molecular biology
ISSN: 1089-8638
Titre abrégé: J Mol Biol
Pays: Netherlands
ID NLM: 2985088R
Informations de publication
Date de publication:
20 11 2020
20 11 2020
Historique:
received:
16
07
2020
revised:
08
10
2020
accepted:
09
10
2020
pubmed:
18
10
2020
medline:
6
3
2021
entrez:
17
10
2020
Statut:
ppublish
Résumé
The αβ-tubulin heterodimer is the fundamental building block of microtubules, making it central to several cellular processes. Despite the apparent simplicity of heterodimerisation, the associated energetics and kinetics remain disputed, largely due to experimental challenges associated with quantifying affinities in the <µM range. We use mass photometry to observe tubulin monomers and heterodimers in solution simultaneously, thereby quantifying the αβ-tubulin dissociation constant (8.48 ± 1.22 nM) and its tightening in the presence of GTP (3.69 ± 0.65 nM), at a dissociation rate >10
Identifiants
pubmed: 33068635
pii: S0022-2836(20)30590-8
doi: 10.1016/j.jmb.2020.10.013
pmc: PMC7763485
pii:
doi:
Substances chimiques
Tubulin
0
Guanosine Triphosphate
86-01-1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6168-6172Subventions
Organisme : Cancer Research UK
ID : FC001163
Pays : United Kingdom
Organisme : Medical Research Council
ID : FC001163
Pays : United Kingdom
Organisme : Wellcome Trust
ID : FC001163
Pays : United Kingdom
Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest Philipp Kukura is a founder, director and shareholder in Refeyn Ltd. Adam Fineberg and Thomas Surrey declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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