The Amino Acid Sequence in Fibrin Responsible for High Affinity Thrombin Binding
The Amino Acid Sequence in Fibrin Responsible for High Affinity Thrombin Binding
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纤维蛋白中负责高亲和力凝血酶结合的氨基酸序列
DOI：
10.1055/s-0037-1615607
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发表时间：
2001
影响因子：
6.7
通讯作者：
M. Mosesson
中科院分区：
文献类型：
--
作者：
D. Meh;K. Siebenlist;S. Brennan;Trudy Holyst;M. Mosesson
人纤维蛋白在其E结构域中具有低亲和力凝血酶结合位点，而在其变体链的羧基末端区域中具有高亲和力结合位点（′ 408 -427）。将该氨基酸序列（VRPEHPAETEYDSLYPEG 1）与已知与凝血酶外切位点结合的其他蛋白质序列（如GPIb、血小板凝血酶受体、血栓调节蛋白和水蛭素中的那些）进行比较，表明没有同源性或共有序列，但Glu和Asp富集是所有序列共有的。这些序列中的酪氨酸硫酸化增强凝血酶外位点结合，但这尚未得到统一的研究。通过电喷雾电离质谱法测定的纤维蛋白原链质量为50，549 Da，比其氨基酸/碳水化合物序列预测的值高151 Da。由于每个硫酸根基团使质量增加80 Da，这表明418和422处的酪氨酸都被硫酸化。制备了一系列重叠肽，用于评价它们对125 I标记的PACK-凝血酶与纤维蛋白结合的抑制。414-427是与408-427一样有效的抑制剂，其结合亲和力依赖于所有羧基末端残基。通过用不可硫酸化的Phe取代' 418或422处的Tyr来制备单Tyr硫酸化肽。与非硫酸化肽相比，在任一Tyr残基处的硫酸化增加了结合竞争，但不如具有4至8倍亲和力的双硫酸化肽有效。反向肽或具有重新定位的Tyr残基的正向序列不能很好地竞争凝血酶结合，表明带电残基的位置对凝血酶结合亲和力是重要的
Summary Human fibrin has a low affinity thrombin binding site in its E domain and a high affinity binding site in the carboxy-terminal region of its variant ’ chain (’408-427). Comparison of the ’ amino acid sequence (VRPEHPAETEYDSLYPEDDL) with other protein sequences known to bind to thrombin exosites such as those in GPIb , the platelet thrombin receptor, thrombomodulin, and hirudin suggests no homology or consensus sequences, but Glu and Asp enrichment are common to all. Tyrosine sulfation in these sequences enhances thrombin exosite binding, but this has not been uniformly investigated. The fibrinogen ’ chain mass determined by electrospray ionization mass spectrometry, was 50,549 Da, a value 151 Da greater than predicted from its amino acid/carbohydrate sequence. Since each sulfate group increases mass by 80 Da, this indicates that both tyrosines at 418 and 422 are sulfated. A series of overlapping ’ peptides was prepared for evaluation of their inhibition of 125I-labeled PPACK-thrombin binding to fibrin. ’414-427 was as effective an inhibitor as ’408-427 and its binding affinity was dependent on all carboxy-terminal residues. Mono Tyr-sulfated peptides were prepared by substituting non-sulfatable Phe for Tyr at ’ 418 or 422. Sulfation at either Tyr residue increased binding competition compared with non-sulfated peptides, but was less effective than doubly sulfated peptides, which had 4 to 8-fold greater affinity. The reverse ’ peptide or the forward sequence with repositioned Tyr residues did not compete well for thrombin binding, indicating that the positions of charged residues are important for thrombin binding affinity
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纤维蛋白血栓和可溶性血浆衍生物的凝血酶活性。
DOI：
--
发表时间：
1983
期刊：
The Journal of laboratory and clinical medicine
影响因子：
--
作者：
Francis,CW;MarkhamJr,RE;Barlow,GH;Florack,TM;Dobrzynski,DM;Marder,VJ
通讯作者：
Marder,VJ
DOI：
10.1073/pnas.96.20.11189
发表时间：
1999-09-28
影响因子：
11.1
作者：
Andersen, H;Greenberg, DL;Davie, EW
通讯作者：
Davie, EW
水蛭素的 COOH 末端结构域。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Naski,MC;Fenton2nd,JW;Maraganore,JM;Olson,ST;Shafer,JA
通讯作者：
Shafer,JA
肝素辅因子 II 的 N 末端酸性结构域在糖胺聚糖存在的情况下介导 α-凝血酶的抑制。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
VanDeerlin,VM;Tollefsen,DM
通讯作者：
Tollefsen,DM
血浆因子 XIII 特异性结合含有 γ 链的纤维蛋白原分子。
DOI：
10.1021/bi9606206
发表时间：
1996
期刊：
Biochemistry.
影响因子：
--
作者：
Siebenlist,KR;Meh,DA;Mosesson,MW
通讯作者：
Mosesson,MW