Sedimentation velocity studies of the vinca-alkaloid-induced self-association of tubulin

John J. Correia; Sharon Lobert; Anthony Frankfurter; Coleman A. Boyd; Bojana Vulevic

doi:10.1117/12.274349

22 May 1997 Sedimentation velocity studies of the vinca-alkaloid-induced self-association of tubulin

John J. Correia, Sharon Lobert, Anthony Frankfurter, Coleman A. Boyd, Bojana Vulevic

Proceedings Volume 2985, Ultrasensitive Biochemical Diagnostics II; (1997) https://doi.org/10.1117/12.274349
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

Vinca alkaloids induce tubulin to self-associate into coiled spirals that further align into sheets and macrotubes. The energetics of spiral formation has been studied by sedimentation velocity in a Beckman Optima XLA analytical ultracentrifuge. The analysis process involves conversion of an absorption based sedimentation pattern into a sedimentation distribution, g(s), and determination of mechanism involving isodesmic ligand-medicated or ligand- mediated plus ligand-facilitated self-association. We have compared the vinca alkaloid-induced self-association of porcine brain tubulin in the presence of 50 (mu) M GTP or GDP. For each drug investigated the affinity is shown to be enhanced by GDP and allosterically linked to GDP/GTP, NaCl and divalent cation binding. These allosteric effectors differentially interact with one another. Thus, GDP enhances self-association over GTP by 0.90 kcal/mol, but the enhancement is reduced to 0.35 kcal/mol by increasing NaCl concentration to 150 mM. High salt stimulates spiral formation but it affects GTP-tubulin preferentially over GDP-tubulin. Divalent cations stimulate spiral formation but GDP-tubulin differentiates between Mg⁺², Ca⁺², and Mn⁺², while GTP-tubulin does not. Divalent cations, Ca⁺² and Mn⁺², induce spiral condensation and macrotube formation, but this is also inhibited by high salt. The differential action of these effectors suggests an interpretation of the energetics in terms of a structural model where charge localization, binding domains and conformational interactions within and between the tubulin heterodimers are responsible for the observed allosteric effects.

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John J. Correia, Sharon Lobert, Anthony Frankfurter, Coleman A. Boyd, and Bojana Vulevic "Sedimentation velocity studies of the vinca-alkaloid-induced self-association of tubulin", Proc. SPIE 2985, Ultrasensitive Biochemical Diagnostics II, (22 May 1997); https://doi.org/10.1117/12.274349

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KEYWORDS

Polymers

Proteins

Software development

Brain

Calcium

Data conversion

Manganese

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