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Title: Combining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ

The crystal structure of the N-terminal part of T. thermophilus DnaJ unexpectedly showed an ordered GF domain and guided the design of a construct enabling the first structure determination of a complete DnaJ cochaperone molecule. By combining the crystal structures with spin-labelling EPR and cross-linking in solution, a dynamic view of this flexible molecule was developed. Hsp70 chaperones assist in a large variety of protein-folding processes in the cell. Crucial for these activities is the regulation of Hsp70 by Hsp40 cochaperones. DnaJ, the bacterial homologue of Hsp40, stimulates ATP hydrolysis by DnaK (Hsp70) and thus mediates capture of substrate protein, but is also known to possess chaperone activity of its own. The first structure of a complete functional dimeric DnaJ was determined and the mobility of its individual domains in solution was investigated. Crystal structures of the complete molecular cochaperone DnaJ from Thermus thermophilus comprising the J, GF and C-terminal domains and of the J and GF domains alone showed an ordered GF domain interacting with the J domain. Structure-based EPR spin-labelling studies as well as cross-linking results showed the existence of multiple states of DnaJ in solution with different arrangements of the various domains, which has implications for themore » function of DnaJ.« less
Authors:
 [1] ;  [2] ; ; ; ; ;  [1] ;  [3] ; ;  [1] ;  [2] ; ;  [1]
  1. MPI for Medical Research, Heidelberg (Germany)
  2. Freie Universitat Berlin, Berlin (Germany)
  3. MPI for Molecular Physiology, Dortmund (Germany)
Publication Date:
OSTI Identifier:
22347818
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 69; Journal Issue: Pt 8; Other Information: PMCID: PMC3727329; PMID: 23897477; PUBLISHER-ID: mh5089; OAI: oai:pubmedcentral.nih.gov:3727329; Copyright (c) Barends et al. 2013; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CAPTURE; CROSS-LINKING; CRYSTAL STRUCTURE; CRYSTALLOGRAPHY; CRYSTALS; DESIGN; ELECTRON SPIN RESONANCE; MATHEMATICAL SOLUTIONS; MOBILITY; MOLECULES; PROTEINS; SOLUTIONS; SPIN; SUBSTRATES