Circular dichroism studies on the effects of ethanol on the conformation of alpha1-acid glycoprotein, alpha1-antitrypsin, deoxyribonuclease, pepsinogen, soybean trypsin inhibitor and unfolded ribonucleases.
The effects of 25 to 75 volume-% ethanol on conformation of human serum alpha1-acid glycoprotein, human serum alpha1-antitrypsin, pancreatic deoxyribonuclease I, porcine pepsinogen, the "Kunitz" trypsin inhibitor from soybeans, and oxidized as well as reduced and S-carboxymethylated ribonucleases were tested by the circular dichroism (CD) probe. It was found that 25 volume-% ethanol had a slight effect, whereas 50--75 vol.-% alcohol significantly altered the conformation. The tertiary structure was perturbed and the polypeptide main chain was reorganized into new conformations of higher helix and beta-structure contents than in the native state. Comparison of the various proteins showed that the degree of reorganization depended chiefly on the cross-linking of the main chain by disulfide bridges. While the unfolded ribonucleases were refolded by 25 vol.-% ethanol into ordered conformations, the native ribonuclease and alpha1-antitrypsin was more sensitive to 25 vol.-% ethanol than the conformation of alpha1-acid glycoprotein, pepsinogen, and soybean trypsin inhibitor. Almost complete restoration of the native conformation was achieved by diluting the alcohol-containing solutions with water or by dialysis against water or buffer solutions. However, the renaturation depended on the time of contact with alcohol and on the temperature at which the alcohol-containing solutions were kept.
D002942:Circular Dichroism; D003851:Deoxyribonucleases; D000431:Ethanol / Q000494:pharmacology*; D009961:Orosomucoid; D010435:Pepsinogens; D011487:Protein Conformation / Q000187:drug effects*; D011489:Protein Denaturation; D012260:Ribonucleases; D014360:Trypsin Inhibitor, Kunitz Soybean; D000515:alpha 1-Antitrypsin
D009961:Orosomucoid; D010435:Pepsinogens; D000515:alpha 1-Antitrypsin; D000431:Ethanol; D014360:Trypsin Inhibitor, Kunitz Soybean; D003851:Deoxyribonucleases; D012260:Ribonucleases