Conserved ATPase and luciferase refolding activities between bacteria and yeast Hsp70 chaperones and modulators.
We have reconstituted an ATP-dependent protein folding machinery using purified yeast cytosolic proteins. The S. cerevisiae Hsp70 Ssa1p and the DnaJ homolog Ydj1p refolded denatured firefly luciferase. In E. coli, efficient refolding of luciferase requires the Hsp70 DnaK and two modulators, DnaJ and GrpE, that synergistically stimulate its ATPase activity. Exchanging DnaJ homologs between the S. cerevisiae and E. coli systems revealed that their ability to stimulate Hsp70 ATPase activity was conserved. In contrast, GrpE further stimulated only DnaK's ATPase activity. Efficient refolding of luciferase by Ssa1p and DnaJ, but not by DnaK and Ydj1p, suggests that a compatible Hsp70/DnaJ homolog pair can act as a protein folding machinery.
Levy
E J
EJ
Department of Anatomy and Cell Biology, State University of New York Health Science Center at Brooklyn 11203, USA.
McCarty
J
J
Bukau
B
B
Chirico
W J
WJ
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D001517:Coleoptera / Q000201:enzymology; D004789:Enzyme Activation; D004926:Escherichia coli / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism*; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D018840:HSP70 Heat-Shock Proteins; D008156:Luciferases; D000251:Adenosine Triphosphatases