Interaction of BiP with the J-domain of the Sec63p component of the endoplasmic reticulum protein translocation complex
Interaction of BiP with the J-domain of the Sec63p component of the endoplasmic reticulum protein translocation complex
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DOI：
10.1074/jbc.274.29.20110
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发表时间：
1999-07-16
影响因子：
4.8
通讯作者：
Rapoport, TA
中科院分区：
文献类型：
--
作者：
Misselwitz, E;Staeck, O;Rapoport, TA
ATP酶的Hsp 70家族的蛋白质与其J蛋白伴侣的保守结构域（J结构域）相互作用，以在许多细胞过程中发挥作用。我们已经研究了BiP的相互作用，热休克蛋白70家族成员在内质网的内腔，与J-结构域的Sec 63 p，一个组件的SEC复杂的翻译后蛋白质易位跨内质网膜。在实时固相结合测定中，BiP在需要ATP水解的反应中与固定化的Sec复合物或与J结构域和谷胱甘肽S-转移酶的融合蛋白结合。在最终的复合物中，BiP以ADP形式结合，其肽结合口袋被占据。这种相互作用需要完整的肽结合口袋。我们的实验表明，由J-结构域激活的BiP涉及这些组件之间的瞬时接触，并且在没有生理底物的情况下，J-激活的BiP甚至与J-蛋白本身结合。
Proteins of the Hsp70 family of ATPases interact with a conserved domain of their J-protein partners, the J-domain, to function in numerous cellular processes. We have studied the interaction of BiP, an Hsp70 family member in the lumen of the endoplasmic reticulum, with the J-domain of Sec63p, a component of the Sec complex involved in post-translational protein translocation across the endoplasmic reticulum membrane. In a real-time solid phase binding assay, BiP binds to the immobilized Sec complex or to a fusion protein of the J-domain and glutathione S-transferase in a reaction that requires ATP hydrolysis. In the final complex, BiP is bound in the ADP form with its peptide binding pocket occupied. An intact peptide binding pocket is required for this interaction. Our experiments suggest that the activation of BiP by the J-domain involves a transient contact between these components, and that in the absence of physiological substrates, J-activated BiP binds even to the J-proteins themselves.