Effects of dimethyl sulfoxide, glycerol, and ethylene glycol on secondary structures of cytochrome c and lysozyme as observed by infrared spectroscopy.
Effects of 10-30% (v/v) of dimethyl sulfoxide, glycerol, and ethylene glycol on the H-O-H bending vibration of water and the amide I bands of horse heart cytochrome c and chicken egg white lysozyme in 25 mM sodium phosphate buffer (pH 7.4) were examined at 20 degrees C by Fourier transform infrared spectroscopy. The H-O-H bending mode of water was strongly affected by these cryoprotectant solvents. Increasing the concentration of cryosolvents from 0 to 30% shifts the water bending band maximum from 1645 to about 1650 cm-1. Second-derivative analysis reveals significant changes in conformation-sensitive amide I regions of lysozyme ascribed to alpha-helix (1657 cm-1), turn (1674 cm-1), and unordered (1646 cm-1) structures; each cryosolvent increases the intensity of the 1657 cm-1 band at the expense of bands at 1674 and 1646 cm-1. No changes in spectra deemed significant were observed for cytochrome c under the same conditions. There is no spectral evidence of structural randomization of proteins due to the presence of these cryosolvents. Cryosolvent-induced changes in secondary structure of proteins may result from changes in water structure which, in turn, perturb the structure of the protein and/or from direct interactions between cryosolvent and protein.
Huang
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P
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins 80523, USA.
Dong
A
A
Caughey
W S
WS
D000818:Animals; D002645:Chickens; D003451:Cryoprotective Agents / Q000737:chemistry*; D003574:Cytochrome c Group / Q000737:chemistry*; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D005026:Ethylene Glycols / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry*; D006736:Horses; D009113:Muramidase / Q000737:chemistry*; D009206:Myocardium / Q000201:enzymology; D010063:Ovum / Q000201:enzymology; D011487:Protein Conformation; D013055:Spectrophotometry, Infrared; D017550:Spectroscopy, Fourier Transform Infrared; D014867:Water / Q000737:chemistry
D003451:Cryoprotective Agents; D003574:Cytochrome c Group; D005026:Ethylene Glycols; D014867:Water; D009113:Muramidase; D005990:Glycerol; D004121:Dimethyl Sulfoxide