title: The structure of denatured bovine pancreatic trypsin inhibitor (BPTI).
abstract: In the presence of denaturant and thiol initiator, the native bovine pancreatic trypsin inhibitor (BPTI) denatures by shuffling its native disulfide bonds and converts to a mixture of scrambled isomers. The extent of denaturation is evaluated by the relative yields of the scrambled and native species of BPTI. BPTI is an exceedingly stable molecule and can be effectively denatured only by guanidine thiocyanate (GdmSCN) at concentrations higher than 3-4 M. The denatured BPTI consists of at least eight fractions of scrambled isomers. Their composition varies under increasing concentrations of GdmSCN. In the presence of 6 M GdmSCN, the most predominant fraction of scrambled BPTI accounts for 56% of the total structure of denatured BPTI. Structural analysis reveals that this predominant fraction contains the bead-form isomer of scrambled BPTI, bridged by three pairs of neighboring cysteines, Cys5-Cys14, Cys30-Cys38 and Cys51-Cys55. The extreme conformational stability of BPTI has important implications in its distinctive folding pathway.
authors.affiliation: Research Center for Protein Chemistry, Institute of Molecular Medicine, The University of Texas, 2121 W. Holcombe Blvd., Houston, TX, USA. rchang@imm2.imm.uth.tmc.edu
authors.forename: J
authors.initials: J
authors.lastname: Chang
authors.forename: A
authors.initials: A
authors.lastname: Ballatore
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000737:chemistry* / Q000187:drug effects; D002417:Cattle; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D019791:Guanidine / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D011487:Protein Conformation; D011489:Protein Denaturation / Q000187:drug effects; D020539:Sequence Analysis, Protein; D013861:Thiocyanates / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
chemical_list: D004220:Disulfides; D006146:Guanidines; D013861:Thiocyanates; C054436:guanidine thiocyanate; D014508:Urea; D007611:Aprotinin; D019791:Guanidine