Author/Editor     Žerovnik, Eva; Jerala, Roman; Kroon-Žitko, Louisa; Pain, Roger H; Turk, Vito
Title     Intermediates in denaturation of a small globular protein, recombinant human stefin B
Type     članek
Source     J Biol Chem
Vol. and No.     Letnik 267, št. 13
Publication year     1992
Volume     str. 9041-6
Language     eng
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.
Descriptors     CYSTATINS
CHROMATOGRAPHY, GEL
CIRCULAR DICHROISM
GUANIDINES
HEAT
HYDROGEN-ION CONCENTRATION
PROTEIN DENATURATION
RECOMBINANT PROTEINS
SPECTROPHOTOMETRY, ULTRAVIOLET