Combining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ
| Autor: | Ralf Seidel, Anna Scherer, Richard Brosi, Andrea Steinmetz, Ilme Schlichting, Thorsten Lorenz, Jochen Reinstein, Robert Bittl, Elisabeth Hartmann, Thomas R. M. Barends, Sabine Zimmermann, Robert L. Shoeman, Jessica Eschenbach |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Models
Molecular endocrine system Protein Folding hybrid structure determination Protein Conformation crystal dehydration Crystal structure single-wavelength anomalous diffraction Crystallography X-Ray law.invention Protein structure Methionine Bacterial Proteins Structural Biology ATP hydrolysis law radiation-damage-induced phasing with anomalous scattering Molecule Electron paramagnetic resonance biology Chemistry Thermus thermophilus molecular chaperones Electron Spin Resonance Spectroscopy General Medicine HSP40 Heat-Shock Proteins biology.organism_classification Research Papers Protein Structure Tertiary Crystallography electron paramagnetic resonance Amino Acid Substitution Chaperone (protein) biology.protein Protein folding cross-linking |
| Zdroj: | Acta Crystallographica Section D: Biological Crystallography Acta Crystallographica. Section D: Biological Crystallography (Copenhagen) |
| ISSN: | 1399-0047 |
| Popis: | 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 the function of DnaJ. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text