title: GroE assists refolding of recombinant human pro-urokinase.
abstract: GroE, one of the molecular chaperones, facilitates correct protein folding both in vitro and in vivo. The refolding of recombinant human pro-urokinase, a protein with a high content of disulfide bonds, was used as a model system to illustrate the mechanism of action of GroE. Aggregation of this protein predominates during its in vitro refolding, as indicated by a strong, concentration-dependent increase in light scattering. The addition of GroE and Mg-ATP significantly increases the yield of the active protein. GroE specifically inhibits the aggregation reaction that competes with correct folding, as shown by a strong decrease in the intensity of light scattering. GroEL rapidly binds to unfolded or partially folded pro-urokinase molecules and thus protects them from the aggregation reaction. Interaction with GroES and ATP hydrolysis are required for the release of the polypeptide chain from GroEL and further acquisition of the completely folded, native conformation.
authors.affiliation: Department of Biochemistry, Nanjing University, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Xu
authors.forename: S
authors.initials: S
authors.lastname: Yang
authors.forename: D
authors.initials: D
authors.lastname: Zhu
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D018833:Chaperonins; D004792:Enzyme Precursors / Q000378:metabolism*; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D005343:Fibrinolytic Agents / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008027:Light; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism; D012542:Scattering, Radiation; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004792:Enzyme Precursors; D029968:Escherichia coli Proteins; D005343:Fibrinolytic Agents; C028715:GroE protein, E coli; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D014568:Urokinase-Type Plasminogen Activator; D018833:Chaperonins; C067551:saruplase