Shi Y, Thomas J O
Department of Biochemistry, New York University School of Medicine, New York 10016.
Mol Cell Biol. 1992 May;12(5):2186-92. doi: 10.1128/mcb.12.5.2186-2192.1992.
The 70-kDa heat shock protein hsp70 and its constitutively expressed cognate, hsc70, are abundant proteins implicated in a number of cellular processes. When a permeabilized cell system for examining the transport of proteins into the nucleus is depleted of hsc70 and hsp70, either by affinity chromatography on ATP-agarose or with antibodies against these proteins, nuclear transport activity is lost. Full activity is restored by the addition of HeLa proteins that bind to ATP-agarose. hsc70 and hsp70 are the active factors, since activity is also fully restored by the addition of either recombinant hsc70 or hsp70 which has been bacterially expressed and highly purified. The restoration of activity is saturable. The transport system requires other cytosolic factors as well, including at least one protein that is sensitive to inactivation by N-ethylmaleimide, but neither hsc70 nor hsp70 is the sensitive protein.
70 kDa热休克蛋白hsp70及其组成性表达的同源蛋白hsc70是参与多种细胞过程的丰富蛋白质。当通过ATP-琼脂糖亲和层析或使用针对这些蛋白质的抗体耗尽用于检测蛋白质向细胞核转运的通透细胞系统中的hsc70和hsp70时,核转运活性丧失。通过添加与ATP-琼脂糖结合的HeLa蛋白可恢复全部活性。hsc70和hsp70是活性因子,因为通过添加细菌表达并高度纯化的重组hsc70或hsp70也可完全恢复活性。活性的恢复是可饱和的。转运系统还需要其他胞质因子,包括至少一种对N-乙基马来酰亚胺失活敏感的蛋白质,但hsc70和hsp70都不是敏感蛋白。