Liu F H, Wu S J, Hu S M, Hsiao C D, Wang C
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China.
J Biol Chem. 1999 Nov 26;274(48):34425-32. doi: 10.1074/jbc.274.48.34425.
Using a yeast two-hybrid system with the 70-kDa heat shock cognate protein (hsc70) or its C-terminal 30-kDa domain as baits, we isolated several proteins interacting with hsc70, including Hip/p48 and p60/Hop. Both are known to interact with hsc70. Except for Hip/p48, all of the proteins that we isolated interact with the 30-kDa domain. Moreover, the EEVD motif at the C terminus of the 30-kDa domain appears essential for this interaction. Sequence analysis of these hsc70-interacting proteins reveals that they all contain tetratricopeptide repeats. Using deletion mutants of these proteins, we demonstrated either by two-hybrid or in vitro binding assays that the tetratricopeptide repeat domains in these proteins are necessary and sufficient for mediating the interaction with hsc70.
利用酵母双杂交系统，以70 kDa热休克同源蛋白（hsc70）或其C端30 kDa结构域作为诱饵，我们分离出了几种与hsc70相互作用的蛋白质，包括Hip/p48和p60/Hop。已知这两种蛋白都能与hsc70相互作用。除了Hip/p48外，我们分离出的所有蛋白质都与30 kDa结构域相互作用。此外，30 kDa结构域C端的EEVD基序似乎对这种相互作用至关重要。对这些与hsc70相互作用的蛋白质进行序列分析发现，它们都含有四肽重复序列。利用这些蛋白质的缺失突变体，我们通过双杂交或体外结合试验证明，这些蛋白质中的四肽重复结构域对于介导与hsc70的相互作用是必要且充分的。
