title: Refolding and reassembly of active chaperonin GroEL after denaturation.
abstract: Conditions are reported that, for the first time, permit the folding and assembly of active chaperonin, GroEL, following denaturation in 8 m urea. The folding could be achieved by dilution or dialysis, and the best yields required the simultaneous presence of ammonium sulfate and the Mg2+ complexes of ATP or ADP. Ammonium sulfate was the key to this particular protocol, since there was a small recovery of oligomer in its presence, but no detectable recovery was induced by ATP or ADP without ammonium sulfate. The refolded/reassembled GroEL could arrest the spontaneous folding of rhodanese, and it could participate in the chaperonin-assisted refolding of rhodanese as effectively as GroEL that had never been unfolded. The results demonstrate that the primary sequence of GroEL contains the information required for its folding, assembly, and function. Thus, in contrast to previous studies, although chaperonins may facilitate GroEL folding, they are not necessary for the acquisition of the functional oligomeric state of this chaperone. This ability to fold denatured GroEL in vitro will facilitate studies of the influences that determine the interesting folding pattern adopted by the native protein.
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760, USA.
authors.forename: J
authors.initials: J
authors.lastname: Ybarra
authors.forename: P M
authors.initials: PM
authors.lastname: Horowitz
mesh_terms: D000645:Ammonium Sulfate; D000818:Animals; D002417:Cattle; D018834:Chaperonin 60 / Q000737:chemistry*; D004926:Escherichia coli; D008274:Magnesium / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D011489:Protein Denaturation; D017510:Protein Folding; D013884:Thiosulfate Sulfurtransferase / Q000737:chemistry; D014508:Urea / Q000737:chemistry
chemical_list: D018834:Chaperonin 60; D009711:Nucleotides; D014508:Urea; D013884:Thiosulfate Sulfurtransferase; D008274:Magnesium; D000645:Ammonium Sulfate