Mechanism of Tubulin Oligomers and Single-Rings Disassembly Catastrophe

Abstract

Cold tubulin dimers coexist with tubulin oligomers and single-rings. These structures are involved in microtubule assembly, however, their dynamics are poorly understood. Using state-of-the-art solution synchrotron time-resolved small-angle X-ray scattering we discovered a disassembly catastrophe (half-life of about 0.1 sec) of tubulin rings and oligomers upon dilution or addition of guanosine triphosphate. A slower disassembly (half-life of about 38 sec) was observed following a temperature increase. Our analysis showed that the assembly and disassembly processes were consistent with an isodesmic mechanism, involving a sequence of reversible reactions at which dimers were rapidly added/removed one at a time, terminated by a two orders-of-magnitude slower ring-closing/opening step. We revealed how assembly conditions varied the mass fraction of tubulin in each of the coexisting structures, the rate constants, and the standard Helmholtz free energies for closing a ring and for longitudinal dimer-dimer associations.