Cloning and characterization of a new soluble murine J-domain protein that stimulates BiP, Hsc70 and DnaK ATPase activity with different efficiencies

Barbara Kroczynska; Sylvie Y. Blond

doi:10.1016/S0378-1119(01)00583-2

Cloning and characterization of a new soluble murine J-domain protein that stimulates BiP, Hsc70 and DnaK ATPase activity with different efficiencies

Barbara Kroczynska, Sylvie Y. Blond^*

^*Corresponding author for this work

Radiation Oncology

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Hsp70s perform many functions in the cell through their ATPase activity that is stimulated by a genuine partner that contains a highly conserved so called J-domain. Here we report the cloning and characterization of a new J-domain protein named MmDjC7. The complete cDNA encodes a putative soluble 22 kDa protein that contains a conserved J-domain, but lacks the G/F- and C-rich regions found in the bacterial Escherichia coli DnaJ. Northern analysis revealed that mmDjC7 mRNA (0.9 kb) is most abundant in the heart and liver tissues. Recombinant hexahistidine tagged MmDjC7 (25 kDa) was efficiently expressed in E. coli and purified to homogeneity. MmDjC7 stimulates the ATPase activity of murine BiP, Hsc70 and E. coli DnaK, albeit with very different molar ratios that vary from 1:2 (for BiP/MmDjC7) to 1:10 (for DnaK/MmDjC7). MmDjC7 thus appears to be a new J-domain protein that can possibly interact with more than one Hsp70.

Original language	English (US)
Pages (from-to)	267-274
Number of pages	8
Journal	Gene
Volume	273
Issue number	2
DOIs	https://doi.org/10.1016/S0378-1119(01)00583-2
State	Published - Aug 8 2001

Funding

We are grateful to Dr Jan A. Miernyk (Plant Research Unit, USDA, ARS, University of Missouri, Columbia, MO) for supplying the human Hsc70 and to Dr Roger McMacken, John Hopkins University, Baltimore, MD for supplying the pRLM163 DnaK expressing plasmid. We thank LaShaunda King, Michael Berg and Mary Hall for helpful discussion and technical advises. This work was supported by grants from the National Institutes of Health (NIHGM-58107) to S.Y.B.

Keywords

Accessory protein
HSP70
Molecular chaperones
Recombinant protein

ASJC Scopus subject areas

Genetics

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10.1016/S0378-1119(01)00583-2

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title = "Cloning and characterization of a new soluble murine J-domain protein that stimulates BiP, Hsc70 and DnaK ATPase activity with different efficiencies",

abstract = "Hsp70s perform many functions in the cell through their ATPase activity that is stimulated by a genuine partner that contains a highly conserved so called J-domain. Here we report the cloning and characterization of a new J-domain protein named MmDjC7. The complete cDNA encodes a putative soluble 22 kDa protein that contains a conserved J-domain, but lacks the G/F- and C-rich regions found in the bacterial Escherichia coli DnaJ. Northern analysis revealed that mmDjC7 mRNA (0.9 kb) is most abundant in the heart and liver tissues. Recombinant hexahistidine tagged MmDjC7 (25 kDa) was efficiently expressed in E. coli and purified to homogeneity. MmDjC7 stimulates the ATPase activity of murine BiP, Hsc70 and E. coli DnaK, albeit with very different molar ratios that vary from 1:2 (for BiP/MmDjC7) to 1:10 (for DnaK/MmDjC7). MmDjC7 thus appears to be a new J-domain protein that can possibly interact with more than one Hsp70.",

keywords = "Accessory protein, HSP70, Molecular chaperones, Recombinant protein",

author = "Barbara Kroczynska and Blond, {Sylvie Y.}",

note = "Funding Information: We are grateful to Dr Jan A. Miernyk (Plant Research Unit, USDA, ARS, University of Missouri, Columbia, MO) for supplying the human Hsc70 and to Dr Roger McMacken, John Hopkins University, Baltimore, MD for supplying the pRLM163 DnaK expressing plasmid. We thank LaShaunda King, Michael Berg and Mary Hall for helpful discussion and technical advises. This work was supported by grants from the National Institutes of Health (NIHGM-58107) to S.Y.B.",

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T1 - Cloning and characterization of a new soluble murine J-domain protein that stimulates BiP, Hsc70 and DnaK ATPase activity with different efficiencies

AU - Kroczynska, Barbara

AU - Blond, Sylvie Y.

N1 - Funding Information: We are grateful to Dr Jan A. Miernyk (Plant Research Unit, USDA, ARS, University of Missouri, Columbia, MO) for supplying the human Hsc70 and to Dr Roger McMacken, John Hopkins University, Baltimore, MD for supplying the pRLM163 DnaK expressing plasmid. We thank LaShaunda King, Michael Berg and Mary Hall for helpful discussion and technical advises. This work was supported by grants from the National Institutes of Health (NIHGM-58107) to S.Y.B.

PY - 2001/8/8

Y1 - 2001/8/8

N2 - Hsp70s perform many functions in the cell through their ATPase activity that is stimulated by a genuine partner that contains a highly conserved so called J-domain. Here we report the cloning and characterization of a new J-domain protein named MmDjC7. The complete cDNA encodes a putative soluble 22 kDa protein that contains a conserved J-domain, but lacks the G/F- and C-rich regions found in the bacterial Escherichia coli DnaJ. Northern analysis revealed that mmDjC7 mRNA (0.9 kb) is most abundant in the heart and liver tissues. Recombinant hexahistidine tagged MmDjC7 (25 kDa) was efficiently expressed in E. coli and purified to homogeneity. MmDjC7 stimulates the ATPase activity of murine BiP, Hsc70 and E. coli DnaK, albeit with very different molar ratios that vary from 1:2 (for BiP/MmDjC7) to 1:10 (for DnaK/MmDjC7). MmDjC7 thus appears to be a new J-domain protein that can possibly interact with more than one Hsp70.

AB - Hsp70s perform many functions in the cell through their ATPase activity that is stimulated by a genuine partner that contains a highly conserved so called J-domain. Here we report the cloning and characterization of a new J-domain protein named MmDjC7. The complete cDNA encodes a putative soluble 22 kDa protein that contains a conserved J-domain, but lacks the G/F- and C-rich regions found in the bacterial Escherichia coli DnaJ. Northern analysis revealed that mmDjC7 mRNA (0.9 kb) is most abundant in the heart and liver tissues. Recombinant hexahistidine tagged MmDjC7 (25 kDa) was efficiently expressed in E. coli and purified to homogeneity. MmDjC7 stimulates the ATPase activity of murine BiP, Hsc70 and E. coli DnaK, albeit with very different molar ratios that vary from 1:2 (for BiP/MmDjC7) to 1:10 (for DnaK/MmDjC7). MmDjC7 thus appears to be a new J-domain protein that can possibly interact with more than one Hsp70.

KW - Accessory protein

KW - HSP70

KW - Molecular chaperones

KW - Recombinant protein

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JO - Gene

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ER -

Cloning and characterization of a new soluble murine J-domain protein that stimulates BiP, Hsc70 and DnaK ATPase activity with different efficiencies

Abstract

Funding

Keywords

ASJC Scopus subject areas

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