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  Structural plasticity and noncovalent substrate binding in the GroEL apical domain - A study using electrospray ionization mass spectrometry and fluorescence binding studies

Ashcroft, A. E., Brinker, A., Coyle, J. E., Weber, F., Kaiser, M., Moroder, L., et al. (2002). Structural plasticity and noncovalent substrate binding in the GroEL apical domain - A study using electrospray ionization mass spectrometry and fluorescence binding studies. Journal of Biological Chemistry, 277(36), 33115-33126.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-6E56-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-6E57-C
Genre: Journal Article
Alternative Title : J. Biol. Chem.

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 Creators:
Ashcroft, A. E., Author
Brinker, A.1, Author              
Coyle, J. E., Author
Weber, F.2, Author              
Kaiser, M.3, Author              
Moroder, L.3, Author              
Parsons, M. R., Author
Jager, J., Author
Hartl, F. U.1, Author              
Hayer-Hartl, M.4, Author              
Radford, S. E., Author
Affiliations:
1Hartl, Franz-Ulrich / Cellular Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565152              
2Former Research Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565145              
3Moroder, Luis / Bioorganic Chemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565160              
4External Organizations, ou_persistent22              

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 Abstract: Advances in understanding how GroEL binds to non-native proteins are reported. Conformational flexibility in the GroEL apical domain, which could account for the variety of substrates that GroEL binds, is illustrated by comparison of several independent crystallographic structures of apical domain constructs that show conformational plasticity in helices H and I. Additionally, ESI-MS indicates that apical domain constructs have co-populated conformations at neutral pH. To assess the ability of different apical domain conformers to bind cochaperone and substrate, model peptides corresponding to the mobile loop of GroES and to helix D from rhodanese were studied. Analysis of apical domain-peptide complexes by ESI-MS indicates that only the folded or partially folded apical domain conformations form complexes that survive gas phase conditions. Fluorescence binding studies show that the apical domain can fully bind both peptides independently. No competition for binding was observed, suggesting the peptides have distinct apical domain-binding sites. Blocking the GroES- apical domain-binding site in GroEL rendered the chaperonin inactive in binding GroES and in assisting the folding of denatured rhodanese, but still capable of binding non-native proteins, supporting the conclusion that GroES and substrate proteins have, at least partially, distinct binding sites even in the intact GroEL tetradecamer.

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Language(s): eng - English
 Dates: 2002-09-06
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 41529
ISI: 000177859000093
 Degree: -

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Title: Journal of Biological Chemistry
  Alternative Title : J. Biol. Chem.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 277 (36) Sequence Number: - Start / End Page: 33115 - 33126 Identifier: ISSN: 0021-9258