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Tubulin polymerization in dimethyl sulfoxide

J Robinson, Y Engelborghs

    The Journal of Biological Chemistry
    |May 25, 1982
    PubMed
    Summary

    Dimethyl sulfoxide (Me2SO) enables tubulin self-assembly into functional microtubules without microtubule-associated proteins (MAPs). This Me2SO-induced assembly is optimal at 8% concentration and stabilizes lateral interactions.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Biophysics

    Background:

    • Tubulin self-assembly into microtubules is crucial for cellular structure and function.
    • Microtubule-associated proteins (MAPs) typically regulate tubulin polymerization.
    • Understanding tubulin assembly independent of MAPs is key to deciphering its fundamental properties.

    Purpose of the Study:

    • To investigate the self-assembly of tubulin in the absence of MAPs using dimethyl sulfoxide (Me2SO).
    • To characterize the properties and optimal conditions for Me2SO-induced microtubule formation.
    • To elucidate the role of Me2SO in stabilizing tubulin polymers.

    Main Methods:

    • Tubulin polymerization assays in MES buffer with varying Me2SO concentrations (6-12% v/v).
    • Characterization of Me2SO-induced polymers using morphology and chemical property analysis.
    • Thermodynamic analysis (Van t'Hoff plot) and kinetic studies of polymerization.
    • Ultracentrifugation to detect oligomer formation.

    Main Results:

    • Tubulin formed polymers resembling microtubules between 6-12% Me2SO.
    • Me2SO microtubules required GTP, were sensitive to cold, Ca2+, colchicine, and pressure.
    • Optimal assembly occurred at 8% Me2SO, with specific thermodynamic parameters (ΔH0 = 26.5 kJ/mol, ΔS0 = 186 J/K·mol).
    • Nucleation kinetics indicated a coefficient of 2, and no oligomers formed without free nucleotide, unlike MAP-containing tubulin.

    Conclusions:

    • Dimethyl sulfoxide (Me2SO) can induce tubulin self-assembly into functional microtubules, mimicking natural ones.
    • Me2SO preferentially stabilizes lateral interactions in tubulin polymers.
    • Nucleation is sensitive to local Me2SO concentration, while propagation is supported by solvent conditions.

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