期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 295, 期 24, 页码 8302-8324出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.RA119.012372
关键词
chaperone; oxidative stress; glutathionylation; 70-kilodalton heat shock protein (Hsp70); protein structure; cysteine modification; disulfide bond; HspA1A; protein homeostasis
资金
- Chinese Ministry of Science and Technology Grant [2017YFA-0504000]
- National Natural Science Foundation of China [31570780, 31770829, 21673278, 31920103011, 31470747, 91849203, 31570857]
- Beijing Natural Science Foundation [5172026]
- Youth Innovation Promotion Association
- Chinese Academy of Sciences (CAS)
- National Laboratory of Biomacromolecules
- CAS Center of Excellence in Biomacromolecules
Heat shock protein 70 (Hsp70) proteins are a family of ancient and conserved chaperones. Cysteine modifications have been widely detected among different Hsp70 family membersin vivo, but their effects on Hsp70 structure and function are unclear. Here, we treated HeLa cells with diamide, which typically induces disulfide bond formation except in the presence of excess GSH, when glutathionylated cysteines predominate. We show that in these cells, HspA1A (hHsp70) undergoes reversible cysteine modifications, including glutathionylation, potentially at all five cysteine residues.In vitroexperiments revealed that modification of cysteines in the nucleotide-binding domain of hHsp70 is prevented by nucleotide binding but that Cys-574 and Cys-603, located in the C-terminal ?-helical lid of the substrate-binding domain, can undergo glutathionylation in both the presence and absence of nucleotide. We found that glutathionylation of these cysteine residues results in unfolding of the ?-helical lid structure. The unfolded region mimics substrate by binding to and blocking the substrate-binding site, thereby promoting intrinsic ATPase activity and competing with binding of external substrates, including heat shock transcription factor 1 (Hsf1). Thus, post-translational modification can alter the structure and regulate the function of hHsp70.
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