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Characterization of denatured proteins by hydrophobic interaction chromatography: A preliminary study
Author(s) -
Bramanti Emilia,
Ferri Fabrizio,
Sortino Chandra,
Onor Massimo,
Raspi Giorgio,
Venturini Marcella
Publication year - 2003
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.10366
Subject(s) - chemistry , myoglobin , hydrophilic interaction chromatography , globular protein , urea , chromatography , denaturation (fissile materials) , cytochrome c , hydrophobic effect , elution , biophysics , crystallography , biochemistry , high performance liquid chromatography , mitochondrion , nuclear chemistry , biology
In this preliminary study hydrophobic interaction chromatography (HIC) is proposed as a good tool in order to detect conformational changes induced by chemical denaturants in two globular proteins, cytochrome C (Cyt C) and myoglobin (MYO). Alterations in protein structure were manifested chromatographically by reproducible changes in peak heights, retention time, and appearance of multiple peaks. The HIC behavior of the two model proteins denatured by guanidinium thyocyanate (GdmSCN) was investigated, keeping constant various concentrations of urea in the mobile phase in a TSK‐Gel Phenyl‐5PW column (TosoBiosep). Suitable elution conditions provide evidence of the simultaneous presence of two denatured forms in the case of MYO, and sequential different denatured states of Cyt C. © 2003 Wiley Periodicals, Inc. Biopolymers 69: 293–300, 2003