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| Content Provider | PubMed Central |
|---|---|
| Author | Sekhar, Ashok Santiago, Margarita Lam, Hon Nam Lee, Jung Ho Silvia, Cavagnero |
| Copyright Year | 2012 |
| Abstract | Most known proteins have at least one local Hsp70 chaperone binding site. Does this mean that all proteins interact with Hsp70 as they fold? This study makes an initial step to address the above question by examining the interaction of the E.coli Hsp70 chaperone (known as DnaK) and its co-chaperones DnaJ and GrpE with a slow-folding E.coli substrate, RNase HD. Importantly, this protein is a nonobligatory client, and it is able to fold in vitro even in the absence of chaperones. We employ stopped-flow mixing, chromatography, and activity assays to analyze the kinetic perturbations induced by DnaK/DnaJ/GrpE (K/J/E) on the folding of RNase HD. We find that K/J/E slows down RNase HD's apparent folding, consistent with the presence of transient chaperone-substrate interactions. However, kinetic retardation is moderate for this slow-folding client and it is expected to be even smaller for faster-folding substrates. Given that the interaction of folding-competent substrates such as RNase HD with the K/J/E chaperones is relatively short-lived, it does not significantly interfere with the timely production of folded biologically active substrate. The above mode of action is important because it preserves K/J/E bioavailability, enabling this chaperone system to act primarily by assisting the folding of other misfolded and (or) aggregation-prone cellular proteins that are unable to fold independently. When refolding is carried out in the presence of K/J and absence of the nucleotide exchange factor GrpE, some of the substrate population becomes trapped as a chaperone-bound partially unfolded state. |
| Related Links | http://dx.doi.org/10.1002/pro.2087 |
| Starting Page | 1042 |
| File Format | |
| ISSN | 1469896X |
| e-ISSN | 1469896X |
| Journal | Protein Science : A Publication of the Protein Society |
| Issue Number | 7 |
| Volume Number | 21 |
| Language | English |
| Publisher | Wiley Subscription Services, Inc., A Wiley Company |
| Publisher Date | 2012-07-01 |
| Access Restriction | Open |
| Rights Holder | Wiley Subscription Services, Inc., A Wiley Company |
| Subject Keyword | Proteomics Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Molecular Biology Biochemistry |
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