title: The pro region of BPTI facilitates folding.
abstract: The in vitro folding pathway of bovine pancreatic trypsin inhibitor (BPTI) has been described previously in terms of the disulfide-bonded intermediates that accumulate during folding of the protein. Folding is slow, occurring in hours at pH 7.3, 25 degrees C. In addition, approximately half of the BPTI molecules become trapped as a dead-end, native-like intermediate. In vivo, BPTI is synthesized as a precursor protein that includes a 13 residue amino-terminal pro region. This pro region contains a cysteine residue. We find that, in vitro, both the rate of formation and the yield of properly folded BPTI are increased substantially in a recombinant model of pro-BPTI. The cysteine residue is necessary for this effect. Moreover, a single cysteine residue, tethered to the carboxy-terminal end of BPTI with a flexible linker of repeating Ser-Gly-Gly residues, is sufficient to assist in disulfide formation. Thus, the pro region appears to facilitate folding by providing a tethered, solvent-accessible, intramolecular thiol-disulfide reagent.
authors.affiliation: Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge 02142.
authors.forename: J S
authors.initials: JS
authors.lastname: Weissman
authors.forename: P S
authors.initials: PS
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000737:chemistry / Q000648:ultrastructure*; D002417:Cattle; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005978:Glutathione / Q000737:chemistry; D066298:In Vitro Techniques; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry; D011498:Protein Precursors / Q000648:ultrastructure*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D004220:Disulfides; D010455:Peptides; D011498:Protein Precursors; D011994:Recombinant Proteins; D007611:Aprotinin; D005978:Glutathione; D003545:Cysteine