title: Intermediates in denaturation of a small globular protein, recombinant human stefin B.
abstract: Guanidinium HCl (GdmHCl), pH, and heat denaturation of the recombinant human stefin B, a low molecular weight protein inhibitor of cysteine proteinases, has been followed by circular dichroism. From the noncoincidence of the transitions in the near and far UV, the existence of stable intermediate states possessing few persistent tertiary interactions but most of the native-like secondary structure, was inferred. These intermediate states exist at equilibrium under various conditions, namely, state G at 1.7 M GdmHCl (pH 8, 25 degrees C), state A at pH 4 (0.6 M GdmHCl, 25 degrees C) and state T above 68 degrees C. By size exclusion chromatography, their apparent compactness was determined. The intermediate states A, T, and G were compact and are therefore classified as "molten globule" states.
authors.affiliation: Department of Biochemistry, Institute J. Stefan, University of Ljubljana, Slovenia.
authors.forename: E
authors.initials: E
authors.lastname: Zerovnik
authors.forename: R
authors.initials: R
authors.lastname: Jerala
authors.forename: L
authors.initials: L
authors.lastname: Kroon-Zitko
authors.forename: R H
authors.initials: RH
authors.lastname: Pain
authors.forename: V
authors.initials: V
authors.lastname: Turk
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D055313:Cystatin B; D015891:Cystatins / Q000737:chemistry*; D019791:Guanidine; D006146:Guanidines; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
chemical_list: C524994:CSTB protein, human; D015891:Cystatins; D006146:Guanidines; D011994:Recombinant Proteins; D055313:Cystatin B; D019791:Guanidine