The 65-kDa cytosolic protein associated with warm temperature acclimation in goldfish,Carassius auratus |
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Authors: | K Kikuchi S Watabe Y Suzuki K Aida H Nakajima |
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Institution: | (1) Laboratory of Fish Physiology, Faculty of Agriculture, The University of Tokyo, 113 Bunkyo, Tokyo, Japan;(2) Laboratory of Marine Biochemistry, Faculty of Agriculture, The University of Tokyo, 113 Bunkyo, Tokyo, Japan;(3) Central Research Institute, Taiyo Fishery Co., Ltd., 300-42 Tsukuba, Ibaraki, Japan |
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Abstract: | Goldfish Carassius auratus were acclimated to either 10 or 30°C for a minimum of 5 weeks. A 65-kDa protein specific to warm-temperature-acclimated fish was extracted from the gel with 70% formic acid after two-dimensional electrophoresis of the muscle cytoplasmic protein fraction. The 65-kDa protein thus prepared to homogeneity was used to raise specific antibodies in rabbit by conventional methods. The antibody produced exhibited specific reaction with a protein having the same molecular weight from brain and liver tissue, suggesting that the 65-kDa protein is a ubiquitous cytosolic component in warm-acclimated goldfish. When water temperature was increased from 20 to 30°C over a 20-h period, a prominent amount of the 65-kDa protein was observed in muscle tissue extracts within 5 days of additional rearing; this was demonstrated by immunoblotting with the specific antibody. The N-terminal amino acid sequence of the 65-kDa protein was determined as Asp-Glu-Pro-Gln-Gly-His-Gln-His (or Asp)-Glu-Leu, differing from that of a family of known heat-shock proteins having about 70 kDa in molecular mass (hsp 70). No interaction between ATP and the 65-kDa protein revealed by ATP-agarose affinity chromatography further confirmed the different properties of the 65-kDa protein from those of hsp 70.Abbreviations ATP
adenosine 5 -triphosphate
- hsp
heat-shock protein(s)
- IgG
immunoglobulin G
- mRNA
messenger ribonucleic acid
- PMSF
phenylmethylsulphonyl fluoride
- PVDF
polyvinylidene difluoride
- SDS
sodium dodecyl sulphate
- SDS-PAGE
SDS-polyacrylamide gel electrophoresis |
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Keywords: | Temperature Acclimation Heat-shock protein Expression Goldfish Carassius auratus |
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