Isolation of a 65-kDa protein from white muscle of warm temperature-acclimated goldfish (Carassius auratus).
A 65-kDa protein expressed in association with warm temperature acclimation of goldfish (Carassius auratus) was purified from epaxial muscle by successive ion-exchange, gel filtration, and reversed-phase columns while monitoring immuno-reaction with a specific antibody. A total of 517 micrograms of the 65-kDa protein was obtained from 23.4 g of the muscle of 30 degrees C-acclimated fish. The purified 65-kDa protein gave one band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was determined to be 65,000 by gel filtration and SDS-PAGE, demonstrating that it consists of a single polypeptide chain; 44 amino acid residues were determined by N-terminal amino acid sequencing. The amino acid stretch was comparatively rich in histidine and phenylalanine. Homology search in the National Biomedical Research Foundation and Swiss-Prot bank did not identify any known amino acid sequence with significant homology to the 44-amino acid stretch of the 65-kDa protein, suggesting it to be a novel protein.
Kikuchi
K
K
Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Watabe
S
S
Aida
K
K
D000064:Acclimatization; D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006054:Goldfish / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007120:Immunochemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D006360:Heat-Shock Proteins; D009124:Muscle Proteins