title: Crystal structures of the 70-kDa heat shock proteins in domain disjoining conformation.
abstract: The 70-kDa heat shock proteins (Hsp70s) are highly conserved ATP-dependent molecular chaperones composed of an N-terminal nucleotide binding domain (NBD) and a C-terminal protein substrate binding domain (SBD) in a bilobate structure. Interdomain communication and nucleotide-dependent structural motions are critical for Hsp70 chaperone functions. Our understanding of these functions remains elusive due to insufficient structural information on intact Hsp70s that represent the different states of the chaperone cycle. We report here the crystal structures of DnaK from Geobacillus kaustophilus HTA426 bound with ADP-Mg(2+)-P(i) at 2.37A and the 70-kDa heat shock cognate protein from Rattus norvegicus bound with ADP-P(i) at 3.5A(.) The NBD and SBD in these structures are significantly separated from each other, and they might depict the ADP-bound conformation. Moreover, a Trp reporter was introduced at the potential interface region between NBD and the interdomain linker of GkDnaK to probe environmental changes. Results from fluorescence measurements support the notion that substrate binding enhances the domain-disjoining behavior of Hsp70 chaperones.
authors.affiliation: Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Chang
authors.forename: Yuh-Ju
authors.initials: YJ
authors.lastname: Sun
authors.forename: Chung
authors.initials: C
authors.lastname: Wang
authors.forename: Chwan-Deng
authors.initials: CD
authors.lastname: Hsiao
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry; D000818:Animals; D001407:Bacillus / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000502:physiology; D011817:Rabbits
chemical_list: D001426:Bacterial Proteins; D018840:HSP70 Heat-Shock Proteins; D000251:Adenosine Triphosphatases