Kampinga H H, Muller E, Brunsting J F, Heine L, Konings A W, Issels R D
Department of Radiobiology, University of Groningen, The Netherlands.
Int J Hyperthermia. 1993 Jan-Feb;9(1):89-98. doi: 10.3109/02656739309061481.
HSP72 levels in the cellular and the nuclear (TX-insoluble) fraction before and after heating of heat- and sodium arsenite-induced thermotolerant and non-tolerant HeLa S3 cells have been investigated by 1D- and 2D-electrophoresis, followed by Western blotting and immunostaining, using a newly developed monoclonal antibody that specifically detects HSP72 (Heine et al. 1991). HSP72 was constitutively expressed in HeLa S3 cells and elevated upon heat or arsenite stress. Immediate association of HSP72 with the nuclear fraction was induced by heat but not arsenite. However, at the time of maximal thermotolerance, elevated levels of HSP72 were found associated with nuclei isolated from both heat- and arsenite-induced thermotolerant cells. After (test) heat treatments (0-60 min at 45 degrees C) translocation of HSP72 to the nuclear fraction in all cells was observed, albeit with different kinetics and to different plateau values. When tolerant and non-tolerant cells were allowed to recover from a heat stress (at 37 degrees C) before isolation of the nuclei, no dissociation of HSP72 from the nuclear fraction was observed within a 5 h time period. Our data indicate that association/dissociation of HSP72 with/from the nuclear fraction is not related to the recovery from heat-induced intranuclear protein aggregation (Kampinga et al. 1992), nor to the extent of thermotolerance in the human HeLa S3 cell line.
利用一种新开发的能特异性检测HSP72的单克隆抗体（Heine等人，1991年），通过一维和二维电泳，随后进行蛋白质免疫印迹和免疫染色，研究了热激和亚砷酸钠诱导的耐热和不耐热HeLa S3细胞在加热前后细胞和细胞核（TX不溶性）组分中的HSP72水平。HSP72在HeLa S3细胞中组成性表达，在热激或亚砷酸钠应激后升高。热激可诱导HSP72立即与细胞核组分结合，而亚砷酸钠则不能。然而，在最大耐热性时，发现从热激和亚砷酸钠诱导的耐热细胞中分离出的细胞核中HSP72水平升高。在（测试）热处理（45℃下0 - 60分钟）后，观察到所有细胞中HSP72向细胞核组分的转位，尽管动力学不同，达到的平台值也不同。当耐热和不耐热细胞在分离细胞核之前从热应激（37℃）中恢复时，在5小时内未观察到HSP72从细胞核组分中解离。我们的数据表明，HSP72与细胞核组分的结合/解离与热诱导的核内蛋白质聚集的恢复无关（Kampinga等人，1992年），也与人类HeLa S3细胞系中的耐热程度无关。
