Relating Molecular Flexibility to Function: A Case Study of Tubulin

• Published in: Biophysical journal Vol. 83; no. 2; pp. 663 - 680
• Main Authors: Keskin, Ozlem, Durell, Stewart R., Bahar, Ivet, Jernigan, Robert L., Covell, David G.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2002; Biophysical Society
• Subjects: Amino Acid Sequence; Animals; Antineoplastic Agents, Phytogenic - pharmacology; Binding Sites; Databases as Topic; Dimerization; Fluid dynamics; Microtubules - drug effects; Microtubules - metabolism; Models, Molecular; Models, Statistical; Molecular biology; Molecular Sequence Data; Mutation; Paclitaxel - pharmacology; Protein Structure, Secondary; Protein Structure, Tertiary; Proteins; Tubulin - chemistry
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/S0006-3495(02)75199-0

Microtubules (MT), along with a variety of associated motor proteins, are involved in a range of cellular functions including vesicle movement, chromosome segregation, and cell motility. MTs are assemblies of heterodimeric proteins, αβ-tubulins, the structure of which has been determined by electron crystallography of zinc-induced, pacilitaxel-stabilized tubulin sheets. These data provide a basis for examining relationships between structural features and protein function. Here, we study the fluctuation dynamics of the tubulin dimer with the aim of elucidating its functional motions relevant to substrate binding, polymerization/depolymerization and MT assembly. A coarse-grained model, harmonically constrained according to the crystal structure, is used to explore the global dynamics of the dimer. Our results identify six regions of collective motion, comprised of structurally close but discontinuous sequence fragments, observed only in the dimeric form, dimerization being a prerequisite for domain identification. Boundaries between regions of collective motions appear to act as linkages, found primarily within secondary-structure elements that lack sequence conservation, but are located at minima in the fluctuation curve, at positions of hydrophobic residues. Residue fluctuations within these domains identify the most mobile regions as loops involved in recognition of the adjacent regions. The least mobile regions are associated with nucleotide binding sites where lethal mutations occur. The functional coupling of motions between and within regions identifies three global motions: torsional and wobbling movements, en bloc, between the α- and β-tubulin monomers, and stretching longitudinally. Further analysis finds the antitumor drug pacilitaxel (TaxotereR) to reduce flexibility in the M loop of the β-tubulin monomer; an effect that may contribute to tightening lateral interactions between protofilaments assembled into MTs. Our analysis provides insights into relationships between intramolecular tubulin movements of MT organization and function.