Brain and egg tubulins from antarctic fishes are functionally and structurally distinct.
The multitubulin hypothesis proposes that chemically distinct tubulins may possess different polymerization properties or may form functionally different microtubules. To test this hypothesis, we have examined the functional properties and the structures of singlet-specific nonneural and neural tubulins from Antarctic fishes. Tubulins were purified from eggs of Notothenia coriiceps neglecta, and from brain tissues of N. coriiceps neglecta or N. gibberifrons, by DEAE ion-exchange chromatography and cycles of microtubule assembly/disassembly. At temperatures between 0 and 20 degrees C, each of these tubulins polymerized efficiently in vitro to yield microtubules of normal morphology. Critical concentrations for polymerization of egg tubulin ranged from 0.057 mg/ml at 3 degrees C to 0.002 mg/ml at 18 degrees C, whereas those for brain tubulin at like temperatures were 4-10-fold larger. Polymerization of both tubulins was entropically driven, but the apparent standard enthalpy and entropy changes for microtubule elongation by egg tubulin (delta Happ0 = +33.9 kcal/mol, delta Sapp0 = +151 entropy units) were significantly greater than values observed for brain tubulin (delta Happ0 = +26.5 kcal/mol, delta Sapp0 = +121 entropy units). Egg tubulin was composed of approximately six alpha and two beta chains and lacked the beta III isotype, whereas brain tubulin was more complex (greater than or equal to 10 of each chain type). Furthermore, egg alpha tubulins were more basic, and their carboxyl termini more resistant to cleavage by subtilisin, than were the alpha chains of brain. We conclude that brain and egg tubulins from the Antarctic fishes are functionally distinct in vitro, due either to qualitative or quantitative differences in isotypic composition, to differential posttranslational modification of shared isotypes, or to both.
Detrich
H W
HW
Department of Biology, Northeastern University, Boston, Massachusetts 02115.
Fitzgerald
T J
TJ
Dinsmore
J H
JH
Marchese-Ragona
S P
SP
D000818:Animals; D000864:Antarctic Regions; D001923:Brain Chemistry; D002417:Cattle; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes; D007525:Isoelectric Focusing; D008854:Microscopy, Electron; D008870:Microtubules / Q000648:ultrastructure; D010063:Ovum / Q000737:chemistry*; D010449:Peptide Mapping; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D013381:Subtilisins / Q000378:metabolism; D013816:Thermodynamics; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*
D014404:Tubulin; D013381:Subtilisins; D003488:Cyanogen Bromide