Fineberg, AdamSurrey, ThomasKukura, Philipp2020-11-232020-11-232020Fineberg A, Surrey T, Kukura P. Quantifying the monomer-dimer equilibrium of tubulin with mass photometry. J Mol Biol. 2020 Nov 20;432(23):6168-72. DOI: 10.1016/j.jmb.2020.10.0130022-2836http://hdl.handle.net/10230/45847The 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 >10-2 s-1. Our results demonstrate the capabilities of mass photometry for quantifying protein-protein interactions and clarify the energetics and kinetics of tubulin heterodimerisation.application/pdfeng© 2020 The Author(s). Published by Elsevier Ltd. This is an open access article distributed under the terms of the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.info:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.jmb.2020.10.013Binding affinityMass photometrySingle moleculeTubulininfo:eu-repo/semantics/openAccess