Cross-linking site in fibrinogen for alpha 2-plasmin inhibitor.
Cross-linking site in fibrinogen for alpha 2-plasmin inhibitor.
复制标题
纤维蛋白原中 α2-纤溶酶抑制剂的交联位点。
DOI：
10.1016/s0021-9258(18)66755-9
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
N. Aoki
中科院分区：
文献类型：
--
作者：
S. Kimura;N. Aoki
一种血浆蛋白酶抑制剂，α 2-纤溶酶抑制剂（α 2 PI），通过活化的凝血因子XIII（血浆转氨酶）与纤维蛋白（原）的α链交联。α 2 PI在交联反应中仅作为活化因子XIII的谷氨酰胺底物（胺受体），并且交联发生在α 2 PI分子的Gln-2和纤维蛋白（原）α链的赖氨酸残基（胺供体）之间，研究了其位置。α 2 PI和纤维蛋白原与活化的凝血因子XIII反应。分离得到的α 2 PI纤维蛋白原A α链复合物，并进行两个循环的Edman降解，第一个循环使用异硫氰酸苯酯，第二个循环使用二甲基氨基偶氮苯-异硫氰酸酯。在第二个循环的裂解阶段后，对含有与A α链交联的二甲基氨基偶氮苯-硫代乙内酰脲-Gln的水相进行CNBr片段化和胰蛋白酶消化。仅发现一种肽在420 nm处具有吸光度峰，表明该肽中存在二甲基氨基偶氮苯-硫代乙内酰脲-Gln。通过分析NH 2-末端氨基酸序列和氨基酸组成，将该肽鉴定为对应于A α链的残基Asn-290-Arg-348。该肽在位置303处含有单个赖氨酸，表明纤维蛋白原A α链的Lys-303是与α 2 PI的Gln-2形成交联的赖氨酸残基。
A plasma proteinase inhibitor, alpha 2-plasmin inhibitor (alpha 2PI), is cross-linked with alpha chain of fibrin(ogen) by activated coagulation Factor XIII (plasma transglutaminase). alpha 2PI serves only as a glutamine substrate (amine acceptor) for activated Factor XIII in the cross-linking reaction, and the cross-linking occurs between Gln-2 of the alpha 2PI molecule and a lysine residue (amine donor) of fibrin(ogen) alpha chain, whose position was investigated. alpha 2PI and fibrinogen were reacted by activated Factor XIII. The resulting alpha 2PI fibrinogen A alpha chain complex was separated and subjected to two cycles of Edman degradation using phenyl isothiocyanate for the first cycle and dimethylaminoazobenzene-isothiocyanate for the second cycle. The aqueous phase after the cleavage stage of the second cycle, containing dimethylaminoazobenzene-thiohydantoin-Gln cross-linked with A alpha chain, was subjected to CNBr fragmentation and tryptic digestion. Only one of the peptides was found to have the peak of absorbance at 420 nm, indicating the presence of dimethylaminoazobenzene-thiohydantoin-Gln in that peptide. The peptide was identified as corresponding to residues Asn-290-Arg-348 of A alpha chain by analyses of the NH2-terminal amino acid sequence and amino acid composition. The peptide contains a single lysine at position 303, indicating that Lys-303 of fibrinogen A alpha chain is the lysine residue that forms a cross-link with Gln-2 of alpha 2PI.