ANALYSIS OF DISULFIDE-CONTAINING FRAGMENTS OF NA+, K+-ATPASE .1. QUANTITATION OF DISULFIDE BONDS OF THE ALPHA-SUBUNIT

被引:0
作者
GEVONDYAN, NM [1 ]
GAVRILYEVA, EE [1 ]
GEVONDYAN, VS [1 ]
MODYANOV, NN [1 ]
机构
[1] KOVALENKO RES INST EXPTL VET,MOSCOW,RUSSIA
来源
BIOLOGICHESKIE MEMBRANY | 1993年 / 10卷 / 06期
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D O I
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中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Ammetric titration with silver nitrate revealed the presence in pig kidney Na+, K+-ATPase of five disulfide bonds and twenty free cysteine residues, most of which are masked. Complete alkylation of all of free SH groups was found possible only after preliminary digestion of the membrane-bound Na+, K+-ATPase. A fraction of disulfide-containing peptides involving three fragments of the alpha-subunit polypeptide chain, namely: Cys(452)-Lys(461), Ile(507)-Lys(519) Val(545)-Phe(558), has been isolated from the tryptic digest alkylated with 4-vinylpyridine. Reduction of SS bonds with beta-mercaptoethanol and alkylation of the released cysteine residues with radiolabeled iodoacetic acid indicated that three above fragments contained cysteine residues that are involved in the formation of two disulfide bonds.
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页码:632 / 640
页数:9
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