Affordable Access

deepdyve-link
Publisher Website

Quantifying the Monomer–Dimer Equilibrium of Tubulin with Mass Photometry

Authors
  • Fineberg, Adam1
  • Surrey, Thomas2, 3, 4
  • Kukura, Philipp1
  • 1 Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3QZ, UK
  • 2 The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK
  • 3 Centre for Genomic Regulation (CRG), Barcelona Institute of Science and Technology (BIST), Dr Aiguader 88, 08003 Barcelona, Spain
  • 4 ICREA, Passeig de Lluis Companys 23, 08010 Barcelona, Spain
Type
Published Article
Journal
Journal of Molecular Biology
Publisher
Elsevier
Publication Date
Nov 20, 2020
Volume
432
Issue
23
Pages
6168–6172
Identifiers
DOI: 10.1016/j.jmb.2020.10.013
PMID: 33068635
PMCID: PMC7763485
Source
PubMed Central
Keywords
Disciplines
  • Brevia
License
Unknown

Abstract

The α β -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 α β -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 >10−2 s−1. Our results demonstrate the capabilities of mass photometry for quantifying protein–protein interactions and clarify the energetics and kinetics of tubulin heterodimerisation.

Report this publication

Statistics

Seen <100 times