title: Polymerization of Antarctic fish tubulins at low temperatures: energetic aspects.
abstract: Tubulins were purified from the brain tissues of three Antarctic fishes, Notothenia gibberifrons, Notothenia coriiceps neglecta, and Chaenocephalus aceratus, by ion-exchange chromatography and one cycle of temperature-dependent microtubule assembly and disassembly in vitro, and the functional properties of the protein were examined. The preparations contained the alpha- and beta-tubulins and were free of microtubule-associated proteins. At temperatures between 0 and 24 degrees C, the purified tubulins polymerized readily and reversibly to yield both microtubules and microtubule polymorphs (e.g., "hooked" microtubules and protofilament sheets). Critical concentrations for polymerization of the tubulins ranged from 0.87 mg/mL at 0 degrees C to 0.02 mg/mL at 18 degrees C. The van't Hoff plot of the apparent equilibrium constant for microtubule elongation at temperatures between 0 and 18 degrees C was linear and gave a standard enthalpy change (delta H degree) of +26.9 kcal/mol and a standard entropy change (delta S degree) of +123 eu. At 10 degrees C, tubulin from N. gibberifrons polymerized efficiently at high ionic strength; the critical concentration increased monotonically from 0.041 to 0.34 mg/mL as the concentration of NaCl added to the assembly buffer was increased from 0 to 0.4 M. Together, the results indicate that the polymerization of tubulins from the Antarctic fishes is entropically driven and suggest that an increased reliance on hydrophobic interactions underlies the energetics of microtubule formation at low temperatures. Thus, evolutionary modification to increase the proportion of hydrophobic interactions (relative to other bond types) at sites of interdimer contact may be one adaptive mechanism that enables the tubulins of cold-living poikilotherms to polymerize efficiently at low temperatures.
authors.affiliation: Department of Biology, Northeastern University, Boston, Massachusetts 02115.
authors.forename: H W
authors.initials: HW
authors.lastname: Detrich
authors.forename: K A
authors.initials: KA
authors.lastname: Johnson
authors.forename: S P
authors.initials: SP
authors.lastname: Marchese-Ragona
mesh_terms: D000818:Animals; D000864:Antarctic Regions; D001923:Brain Chemistry; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003080:Cold Temperature; D005399:Fishes / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D015394:Molecular Structure; D009994:Osmolar Concentration; D011108:Polymers; D012492:Salts; D013696:Temperature; D013816:Thermodynamics; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D011108:Polymers; D012492:Salts; D014404:Tubulin