Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein

Flaherty, Kevin; DeLuca-Flaherty, Camille; McKay, David

doi:10.1038/346623a0

Cited by 987 publications

(723 citation statements)

References 39 publications

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“…The crystal structure of Hsp70 shows that it consists of a ~ 44 kDa N-terminal domain (ATPase domain) that mediates ATP binding and hydrolysis (Flaherty et al, 1990) and a ~ 27 kDa C-terminal peptide binding domain (Bukau and Horwich, 1998;Zhu et al, 1996) (Figure 4a). The C-terminal domain contains a β-sandwich subdomain with a peptide binding cleft and a α-helical latch-like segment that acts as a lid to permit the entry and release of the substrate.…”

Section: Ii41121 the Hsp70 Chaperone Systemmentioning

confidence: 99%

Firefly luciferase mutants as sensors of proteome stress

et al. 2011

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Section: Ii41121 the Hsp70 Chaperone Systemmentioning

confidence: 99%

Firefly luciferase mutants as sensors of proteome stress

et al. 2011

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“…The crystal structure of the N-terminal ATPase domain of bovine hsc70 shows this region of the protein to have 38% a-helix and 30% P-strand (Flaherty et al, 1990), and the crystal structure of the peptide binding domain of DnaK reveals 33% a-helix and 27% P-strand in this region (Zhu et al, 1996); the CD results for Hsc66 60 h Far UV Circular Dichroism…”

Section: Hsc66 Hsc2omentioning

confidence: 99%

Hsc66 and Hsc20, a new heat shock cognate molecular chaperone system from Escherichia coli

1997

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The hscA and hscB genes of Escherichia coli encode novel chaperone and co-chaperone proteins, designated Hsc66 and Hsc20, respectively. We have overproduced and purified Hsc66 and Hsc2O in high yield in E. coli and describe their initial characterization including absorbance, fluorescence, and circular dichroism spectra. Immunoblot analyses of E. coli cultures using antisera to Hsc66 and Hsc20 raised in rabbits establish that Hsc66 and Hsc2O are constitutively expressed at levels corresponding to cell concentrations -20 pM and -10 pM, respectively. The levels do not change appreciably following heat shock (44"C), but a small increase in Hsc2O is observed following a shift to 10°C. Purified Hsc66 exhibits a low intrinsic ATPase activity (-0.6 min" at 37"C), and Hsc20 was found to stimulate this activity up to 3.8-fold with half-maximal stimulation at a concentration -5 pM. These findings suggest that Hsc66 and Hsc20 comprise a molecular chaperone system similar to the prokaryotic DnaK/Dnd and eukaryotic hsp70/hsp40 systems. Sequence differences between Hsc66 and Hsc20 compared to other members of this chaperone family, however, suggest that the Hsc66/Hsc20 system will display different peptide binding specificity and that it is likely to be subject to different regulatory mechanisms. The high level of constitutive expression and the lack of a major response to temperature changes suggest that Hsc66 and Hsc20 play an important cellular role(s) under non-stress conditions. Keywords: ATPase activity; chaperone; co-chaperone; constitutive expression; hsp70; hsp40; purification The hsp70, or stress-70, proteins comprise a large and widespread family of proteins -7O-kDa, which function as ATP-dependent molecular chaperones (reviewed in Gething

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“…HSP70s belong to one of the most conserved protein families [5]. The characteristic features of a HSP70 protein are a conserved amino (N)-terminal ATPase domain of approximately 44-kDa [6] and a carboxyl (C)-terminal peptide binding domain of approximately 25-kDa which is further subdivided into a b-sandwich subdomain of 15 kDa and a C-terminal a-helical subdomain [7]. HSP70s function as molecular chaperones in the folding and refolding of nonnative proteins to prevent irreversible aggregation [8][9][10], and play roles in protein transport and assembly processes [11,12].…”

Section: Introductionmentioning

confidence: 99%

Characterization of a wheat HSP70 gene and its expression in response to stripe rust infection and abiotic stresses

et al. 2010

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Members of the family of 70-kD heat shock proteins (HSP70 s) play various stress-protective roles in plants. In this study, a wheat HSP70 gene was isolated from a suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The gene, that was designated as TaHSC70, was predicted to encode a protein of 690 amino acids, with a molecular mass of 73.54 KDa and a pI of 5.01. Further analysis revealed the presence of a conserved signature that is characteristic for HSP70s and phylogenetic analysis demonstrated that TaHSC70 is a homolog of chloroplast HSP70s. TaHSC70 mRNA was present in leaves of both green and etiolated wheat seedlings and in stems and roots. The transcript level in roots was approximately threefold less than in leaves but light-dark treatment did not charge TaHSC70 expression. Following heat shock of wheat seedlings at 40°C, TaHSC70 expression increased in leaves of etiolated seedlings but remained stable at the same level in green seedlings. In addition, TaHSC70 was differentially expressed during an incompatible and compatible interaction with wheat-stripe rust, and there was a transient increase in expression upon treatment with methyl jasmonate (MeJA) treatment. Salicylic acid (SA), ethylene (ET) and abscisic acid (ABA) treatments had no influence on TaHSC70 expression. These results suggest that TaHSC70 plays a role in stress-related responses, and in defense responses elicited by infection with stripe rust fungus and does so via a JA-dependent signal transduction pathway.

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Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein

Cited by 987 publications

References 39 publications

Firefly luciferase mutants as sensors of proteome stress

Firefly luciferase mutants as sensors of proteome stress

Hsc66 and Hsc20, a new heat shock cognate molecular chaperone system from Escherichia coli

Characterization of a wheat HSP70 gene and its expression in response to stripe rust infection and abiotic stresses

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