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A gram‐negative characteristic segment in Escherichia coli DnaK is essential for the ATP‐dependent cooperative function with the co‐chaperones DnaJ and GrpE
Author(s) -
Sugimoto Shinya,
Higashi Chihana,
Saruwatari Kozue,
Nakayama Jiro,
Sonomoto Kenji
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.05.055
Subject(s) - escherichia coli , chaperone (clinical) , complementation , bacteria , biology , mutant , atpase , protein fragment complementation assay , gram negative bacteria , biochemistry , enterobacteriaceae , heat shock protein , luciferase , microbiology and biotechnology , enzyme , genetics , transfection , gene , medicine , pathology
We describe importance of the characteristic segment in ATPase domain of DnaK chaperone which is present in all gram‐negative bacteria but is absent in all gram‐positive bacteria. In vitro studies, ATPase activity, luciferase‐refolding activity, and surface plasmon resonance analyses, demonstrated that a segment‐deletion mutant DnaKΔ74‐96 became defective in the cooperation with the co‐chaperones DnaJ and GrpE. In addition, in vivo complementation assay showed that expression of DnaKΔ74‐96 could not rescue the viability of Escherichia coli ΔdnaK mutant at 43 °C. Consequently, we suggest evolutionary significance for this DnaK ATPase domain segment in gram‐negative bacteria towards the DnaK chaperone system.