Kondo Hidehiro, Harano Ryohei, Nakaya Misako, Watabe Shugo
Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 1138657, Japan.
Cell Stress Chaperones. 2004 Winter;9(4):350-8. doi: 10.1379/csc-55r.1.
Temperature-dependent changes of growth rate and protein components were investigated for primary cultured cells derived from goldfish caudal fin. When the culture temperature was shifted from 20 degrees C to 35 degrees C and 40 degrees C, the growth rate was increased at 35 degrees C as compared with that at 20 degrees C, but no cell growth was observed at 40 degrees C. The differential scanning calorimetry demonstrated the onset of the endothermic reaction for goldfish cellular components at 40 degrees C. Therefore, the temperature shift to 40 degrees C was found to be of severe heat shock for goldfish cultured cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that, although expression of 70-kDa components was slightly induced at 35 degrees C, the temperature shift to 40 degrees C markedly induced the expression of the 30-kDa component in addition to that of 70-kDa component. The N-terminal amino acid sequencing identified the 30- and 70-kDa components to be heat shock protein (Hsp)-30 and Hsp70, respectively. Northern blot analysis revealed that the enhanced Hsp30 messenger ribonucleic acid (mRNA) levels were only observed at 40 degrees C, whereas Hsp70 mRNA was slightly accumulated at 35 degrees C. These results indicated that Hsp30 might have important functions under severe heat stress condition.
研究了金鱼尾鳍原代培养细胞生长速率和蛋白质成分随温度的变化。当培养温度从20℃转移到35℃和40℃时，与20℃相比，35℃时生长速率增加，但在40℃未观察到细胞生长。差示扫描量热法表明，金鱼细胞成分在40℃时开始出现吸热反应。因此，发现温度转移到40℃对金鱼培养细胞是严重的热休克。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析显示，虽然在35℃时70-kDa成分的表达略有诱导，但温度转移到40℃除了诱导70-kDa成分的表达外，还显著诱导了30-kDa成分的表达。N端氨基酸测序确定30-kDa和70-kDa成分分别为热休克蛋白（Hsp）-30和Hsp70。Northern印迹分析显示，仅在40℃观察到Hsp30信使核糖核酸（mRNA）水平增强，而Hsp70 mRNA在35℃时略有积累。这些结果表明，Hsp30可能在严重热应激条件下具有重要功能。
