Open AccessCombining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ
Author(s) -
Barends Thomas R. M.,
Brosi Richard W. W.,
Steinmetz Andrea,
Scherer Anna,
Hartmann Elisabeth,
Eschenbach Jessica,
Lorenz Thorsten,
Seidel Ralf,
Shoeman Robert L.,
Zimmermann Sabine,
Bittl Robert,
Schlichting Ilme,
Reinstein Jochen
Publication year - 2013
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s0907444913010640
Subject(s) - thermus thermophilus , chaperone (clinical) , crystallography , atp hydrolysis , site directed spin labeling , protein folding , electron paramagnetic resonance , chemistry , biology , biophysics , biochemistry , escherichia coli , physics , enzyme , gene , atpase , medicine , pathology , nuclear magnetic resonance , membrane
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.