Kinetic analysis of the hydrodynamic transition accompanying protein folding using size exclusion chromatography. 2. Comparison of spectral and chromatographic kinetic analyses.
The kinetics of the hydrodynamic volume change associated with the unfolding and refolding of a globular protein can be observed using high performance size exclusion chromatography. Chromatographic profiles that evidence such dynamics can be simulated using equations in which chromatographic partitioning and the conformational transition are described in terms of a finite difference algorithm incorporating an apparent binding model to generate broad and asymmetric peaks. Application of these equations to the simple two-state unfolding transition of ribonuclease A in guanidine hydrochloride indicates that reliable kinetic parameters can be obtained using these equations.
Shalongo
W
W
Department of Biochemistry, University of Iowa, Iowa City 52242.
Jagannadham
M
M
Stellwagen
E
E
D002850:Chromatography, Gel / Q000379:methods; D004798:Enzymes / Q000737:chemistry; D019791:Guanidine; D006146:Guanidines; D007700:Kinetics; D008433:Mathematics; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D012260:Ribonucleases / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D004798:Enzymes; D006146:Guanidines; D009842:Oligopeptides; D010455:Peptides; D011506:Proteins; D012260:Ribonucleases; D019791:Guanidine