Nimmesgern E, Hartl F U
Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
FEBS Lett. 1993 Sep 27;331(1-2):25-30. doi: 10.1016/0014-5793(93)80290-b.
The protein folding capacity of rabbit reticulocyte cytosol was analyzed using the renaturation of firefly luciferase as a sensitive assay. In the absence of ATP, the aggregation of denatured luciferase diluted into reticulocyte lysate was prevented. Chaperone-stabilized luciferase was detected in high molecular weight complexes overlapping the distributions of Hsc70, Hsp90 and the chaperonin TRiC on gel filtration columns. The readdition of unfractionated cytosol and Mg-ATP was required for the efficient folding of these forms of luciferase to the active enzyme. We conclude that protein folding in the eukaryotic cytosol depends on the functional cooperation of different chaperone activities and cofactors in a complex, ATP-dependent process.
利用萤火虫荧光素酶的复性作为一种灵敏的检测方法，对兔网织红细胞胞质溶胶的蛋白质折叠能力进行了分析。在没有ATP的情况下，稀释到网织红细胞裂解物中的变性荧光素酶的聚集被阻止。在凝胶过滤柱上，在与Hsc70、Hsp90和伴侣蛋白TRiC分布重叠的高分子量复合物中检测到伴侣蛋白稳定的荧光素酶。这些形式的荧光素酶有效折叠成活性酶需要重新添加未分级的胞质溶胶和Mg-ATP。我们得出结论，真核细胞胞质溶胶中的蛋白质折叠取决于不同伴侣蛋白活性和辅因子在一个复杂的、ATP依赖的过程中的功能协作。
