title: GroEL chaperone binding to beetle luciferases and the implications for refolding when co-expressed.
abstract: The folding of many proteins including luciferase in vivo requires the assistance of molecular chaperone proteins. To understand how a chaperone targets luciferase, we took three luciferases that give different bioluminescence with the same luciferin substrate and with differences in homology. The three luciferase genes, firefly luciferase (FF-Luc) (from Pyrocoelia miyako), and red (RE-Luc) and green (GR-Luc) bioluminescence-emitting luciferases (from Phrixothrix railroad-worms), were expressed in Escherichia coli to produce fusion proteins with predicted molecular masses. Subsequently, we observed that DnaK and GroEL were co-purified along with recombinant luciferase. Although the amount of co-purified DnaK was almost the same compared to FF-Luc, GroEL was 25 and 32 times higher in GR-Luc and RE-Luc respectively. Furthermore, co-expression of GroEL/GroES along with luciferase substantially refolded RE-Luc and GR-Luc compared to FF-Luc.
authors.affiliation: Cell Dynamics Research Group, National Institute of Advanced Industrial Science and Technology, Ikeda, Osaka 563-8577, Japan. venkatesh_balan@hotmail.com
authors.forename: Balan
authors.initials: B
authors.lastname: Venkatesh
authors.forename: Mohammad
authors.initials: M
authors.lastname: Arifuzzaman
authors.forename: Hirotada
authors.initials: H
authors.lastname: Mori
authors.forename: Takahisa
authors.initials: T
authors.lastname: Taguchi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmiya
mesh_terms: D000818:Animals; D018834:Chaperonin 60 / Q000378:metabolism*; D001517:Coleoptera / Q000201:enzymology*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D049454:Firefly Luciferin / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D049454:Firefly Luciferin; D008156:Luciferases; C063842:dnaK protein, E coli