Effective cotranslational folding of firefly luciferase without chaperones of the Hsp70 family.
Molecular chaperones of the Hsp70 family (bacterial DnaK, DnaJ, and GrpE) were shown to be strictly required for refolding of firefly luciferase from a denatured state and thus for effective restoration of its activity. At the same time the luciferase was found to be synthesized in an Escherichia coli cell-free translation system in a highly active state in the extract with no chaperone activity. The addition of the chaperones to the extract during translation did not raise the activity of the enzyme. The abrupt arrest of translation by the addition of a translational inhibitor led to immediate cessation of the enzyme activity accumulation, indicating the cotranslational character of luciferase folding. The results presented suggest that the chaperones of the Hsp70 family are not required for effective cotranslational folding of firefly luciferase.
Svetlov
Maxim S
MS
Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia.
Kommer
Aigar
A
Kolb
Vyacheslav A
VA
Spirin
Alexander S
AS
D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry*; D049409:Luciferases, Firefly / Q000737:chemistry* / Q000235:genetics; D018832:Molecular Chaperones / Q000737:chemistry; D014176:Protein Biosynthesis; D017510:Protein Folding; D046188:Protein Modification, Translational
D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D049409:Luciferases, Firefly