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- W3189250091 abstract "Tubulin nucleation, microtubule assembly, stability, and dynamics depend on fluxes of chemical energy, controlled by hydrolysis of guanosine triphosphate (GTP) to guanosine diphosphate (GDP), and nucleotide exchange reactions. In this paper, we determined how tubulin self-association in glycerol-free assembly buffers affects the rate of GTP hydrolysis and the thermodynamics of nucleotide exchange. In the absence of tubulin, GTP hydrolysis was negligible. In the presence of tubulin, below the critical conditions for microtubule assembly, no GTP hydrolysis was observed, even though tubulin 1D curved oligomers and single rings were formed, suggesting that GTP hydrolysis was not involved in their formation. Under conditions permitting spontaneous tubulin nucleation and microtubule assembly, GTP hydrolysis was detected and followed pseudo-first-order kinetics, limited by the rate of tubulin nucleation and microtubule assembly. By simultaneously determining the concentrations of tubulin-free and tubulin-bound GTP and GDP at steady-state, we investigated the nucleotide exchange reaction under conditions where GTP hydrolysis was negligible. The exchange reaction strongly depended on the molar ratio between tubulin-free GDP and GTP and the total tubulin concentration. To analyze these data, we used a thermodynamic model of isodesmic tubulin self-association, terminated by the formation of tubulin single-rings, to calculate the distribution of tubulin single rings, 1D oligomers, and free dimers, and thereby the molar fractions of tubulin dimers with exposed and buried nucleotide exchangeable sites (E-sites). Our data suggest that the exchange reaction occurred to a different extent on tubulin dimers with buried E-sites than on dimers with exposed E-sites." @default.
- W3189250091 created "2021-08-16" @default.
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- W3189250091 date "2021-08-05" @default.
- W3189250091 modified "2023-10-07" @default.
- W3189250091 title "Effect of Tubulin Self-Association on GTP Hydrolysis and Nucleotide Exchange Reactions" @default.
- W3189250091 doi "https://doi.org/10.33774/chemrxiv-2021-71g6t" @default.
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