[Participation of DnaK chaperone and ATP in in vivo folding of firefly luciferase, synthesized by E. coli cells].
The time-courses of protein accumulation and luciferase activity of Luciola mingrelica firefly luciferase synthesized on plasmid pJG lambda in E. coli strains omega 238 and B178groE7 (DnaK- and GroEL-deficient, correspondingly) and omega 237 and W3110 that are control to them, have been studied. It was shown that the amounts of the luciferase protein synthesized by all strains was approximately equal to 3.0-3.9% of the total cell protein. Luciferase was synthesized in a catalytically inactive form and transformed into an active enzyme during incubation at 21 degrees C. In the DnaK-deficient E. coli strain omega 238, the enzyme transformation into a catalytically active form did not occur which provides evidence for participation of the DnaK chaperone in transformation of newly synthesized luciferase into catalytically active form. Luciferase folding was accelerated with an increase in ATP concentration inside the cell. It is possible to increase the ATP concentration inside the cell by treatment with polymyxin and addition of ATP to the culture medium, which significantly accelerates the luciferase folding.
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D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001517:Coleoptera / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D008156:Luciferases; C063842:dnaK protein, E coli
Uchastie shaperona DnaK i ATP v svorachivanii in vivo liutsiferazy svetliako, sinteziruemoĭ kletkami E. coli.