Sequence-specific 1H NMR assignments, secondary structure, and location of the calcium binding site in the first epidermal growth factor like domain of blood coagulation factor IX.
Sequence-specific 1H NMR assignments, secondary structure, and location of the calcium binding site in the first epidermal growth factor like domain of blood coagulation factor IX.
复制标题
凝血因子 IX 的第一个表皮生长因子样结构域中的序列特异性 1H NMR 分配、二级结构和钙结合位点的位置。
DOI：
10.1021/bi00244a006
复制
发表时间：
1991
期刊：
影响因子：
2.9
通讯作者：
Sweeney,WV
中科院分区：
文献类型：
--
作者：
Huang,LH;Cheng,H;Pardi,A;Tam,JP;Sweeney,WV
因子IX是一种凝血蛋白，包含三个区域，包括α-羧基谷氨酸（Gla）结构域、两个串联连接的表皮生长因子样（EGF样）结构域和一个丝氨酸蛋白酶区域。除了Gla结构域中的钙结合之外，该蛋白在第一EGF样结构域中表现出高亲和力的钙结合位点。第一个EGF样结构域，因子IX（45-87），已经合成。用2DNMR技术对该肽进行了序列特异性共振归属，并确定了其二级结构。该蛋白质被发现有两个反平行/3-片层，初步的距离几何计算表明，该蛋白质有两个结构域，由Trp 28分开，与EGF的整体结构相似。钙结合的第一EGF样结构域的NMR研究表明，钙结合位点的存在和位置涉及的两条链上的一个β-片层的残基，以及N-末端区域的肽。这些结果表明，钙结合在第一个EGF样域可以诱导长距离（可能interdomain）的构象变化的因子IX，而不是导致结构改变的EGF样域本身。因子IX是一种维生素K依赖性凝血蛋白质的重要的内在凝血级联。该蛋白具有由N-末端羧基谷氨酸（Gla）1结构域、两个表皮生长因子样（EGF样）结构域和C-末端丝氨酸蛋白酶结构域组成的结构域结构。多种凝血蛋白含有表皮生长因子样结构域的串联重复，包括
Revised Manuscript Received April 2, 1991 abstract: Factor IX is a blood clotting protein that contains three regions, including a-carboxyglutamic acid (Gla) domain, two tandemly connected epidermal growth factor like (EGF-like) domains, and a serine protease region. The protein exhibits a high-affinity calcium binding site in the first EGF-like domain, in addition to calcium binding in the Gla domain. The first EGF-like domain, factor IX (45-87), has been synthesized. Sequence-specific resonance assignment of the peptide has been madeby using 2D NMR techniques, and its secondary structure has been determined. The protein is found to have two antiparallel/3-sheets, and preliminary distance geometry calculations indicate that the protein has two domains, separated by Trp28, with the overall structure being similar to that of EGF. An NMR investigation of the calcium-bound first EGF-likedomain indicates the presence and location of a calcium binding site involving residues on both strands of one of the/3-sheets as well as the N-terminal region of the peptide. These results suggest that calcium binding in the first EGF-likedomain could induce long-range (possibly interdomain) con-formational changes in factor IX, rather than causing structural alterations in the EGF-like domain itself.Factor IX is a vitamin K dependent blood clotting protein important in the intrinsic clotting cascade. This protein has a domain structure consisting of an N-terminal-carboxy-glutamic acid (Gla) 1 domain, two epidermal growth factor like (EGF-like) domains, and a C-terminal serine protease domain. A wide range of blood clotting proteins contain tandem repeats of an epidermal growth factor like domain, including