REDUCTION OF DISULFIDE BONDS IN PROTEINS BY 2-AMINOTHIOPHENOL UNDER WEAKLY ACIDIC CONDITIONS

被引:6
作者
ABE, Y [1 ]
UEDA, T [1 ]
IMOTO, T [1 ]
机构
[1] KYUSHU UNIV 62,FAC PHARMACEUT SCI,HIGASHI KU,FUKUOKA 812,JAPAN
关键词
2-AMINOTHIOPHENOL; LYSOZYME; STI; REDUCTION OF DISULFIDE BOND; RIBONUCLEASE-A;
D O I
10.1093/oxfordjournals.jbchem.a124304
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We developed a method for reducing disulfide bonds in proteins under weakly acidic conditions by use of 2-aminothiophenol. The disulfide bonds in hen egg-white lysozyme, ribonuclease A, and soybean trypsin inhibitor were quantitatively reduced by 2-aminothiophenol in phosphate buffer, pH 6, containing 8 M Gdn HCl, 1 mM EDTA, and 20% ethanol, for 60 min at 40-degrees-C. On analysis of the RP-HPLC patterns of tryptic peptides, which were derived from reduced and S-alkylated lysozyme and ribonuclease A at pH 6, it was confirmed that no side reaction occurred. Moreover, the reduction under weakly acidic conditions was demonstrated to be applicable for the location of such a labile residue as O-acetylated tyrosine.
引用
收藏
页码:52 / 57
页数:6
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