Temperature-induced changes in protein structures studied by Fourier transform infrared spectroscopy and global analysis.
Fourier transform infrared (FTIR) spectroscopy has been used to study temperature-induced structural changes which occur in albumin, immunoglobulin G, fibrinogen, lysozyme, alpha-lactalbumin, and ribonuclease S when dissolved in 2H2O. In order to analyze the data, a new method was developed in which the data were analyzed globally with the aid of a spectral model. Seven or eight bands were sufficient to fit the full data set of spectra ranging from 1420 to 1760 cm-1 with a root mean square error of 1-2% of the maximum. Subsequently, the estimated band amplitude curves which showed a sigmoidal progression with increasing temperature were (globally) fitted with a two-state thermodynamic model. In this way, information on structural changes as well as on the thermal stability of the proteins was obtained. In all proteins investigated, enhanced 1H-2H exchange occurred at temperatures well below the unfolding of the secondary structure. This was interpreted as a change in tertiary structure leading to enhanced solvent accessibility. In all the proteins investigated, except for ribonuclease S, an intermolecular beta-sheet band indicative of aggregation appeared concomitant with the denaturation of the secondary structure. The results are compared with data from other techniques and discussed in terms of local unfolding and folding intermediates.
van Stokkum
I H
IH
Faculty of Physics and Astronomy, Vrije Universiteit, Amsterdam, The Netherlands.
Linsdell
H
H
Hadden
J M
JM
Haris
P I
PI
Chapman
D
D
Bloemendal
M
M
D000818:Animals; D002417:Cattle; D005340:Fibrinogen / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007768:Lactalbumin / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012260:Ribonucleases / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D013816:Thermodynamics
D007074:Immunoglobulin G; D011506:Proteins; D012709:Serum Albumin; D005340:Fibrinogen; D007768:Lactalbumin; D012260:Ribonucleases; C022542:ribonuclease S; D009113:Muramidase