cDNA sequences for human von Willebrand factor reveal five types of repeated domains and five possible protein sequence polymorphisms.
cDNA sequences for human von Willebrand factor reveal five types of repeated domains and five possible protein sequence polymorphisms.
复制标题
人类血管性血友病因子的 cDNA 序列揭示了五种类型的重复结构域和五种可能的蛋白质序列多态性。
DOI：
10.1021/bi00359a014
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Sadler,JE
中科院分区：
文献类型：
--
作者：
Shelton-Inloes,BB;Titani,K;Sadler,JE
霍华德休斯医学研究所实验室，医学和生物化学系，华盛顿大学医学院，圣路易斯，密苏里州63110，以及生物化学系，华盛顿大学，西雅图，华盛顿98195，1986年1月14日接收;修订版Mandarin pt，1986年4月11日接收摘要：用两个先前描述的人血管性血友病因子的cDNA插入片段筛选Xgt 11中的人脐静脉内皮细胞cDNA文库。在16株阳性分离株中，有2株与血管性血友病因子氨基端探针杂交，并进行了序列测定。总之，这四个cDNA插入片段跨越血管性血友病因子mRNA序列的6.5个碱基，完全指定成熟分泌的血管性血友病因子亚基的2050个氨基酸和前体肽的24个残基。大约77%的序列包含在五种类型的重复结构域中。结构域A由193-220个氨基酸组成，并且在残基497和1111之间以三个串联拷贝存在。结构域B含有25-35个氨基酸，并且在残基1533和1636之间以三个拷贝存在。结构域C由116-119个氨基酸组成，并且在残基1637和1899之间重复。与结构域AC的基本上连续的重复相反，结构域D和E的两个拷贝各自分别被804和1383个氨基酸分开。结构域D1在残基79和367之间含有289个氨基酸，而结构域D2在残基1171和1440之间由270个氨基酸组成。结构域E1由残基25和70之间的46个氨基酸组成，结构域E2由残基1453和1498之间的46个氨基酸组成。三重A结构域的Cys含量明显不足，而其余结构域富含Cys。A结构域与补体因子B的225个残基片段同源。另外，血管性血友病因子与国家生物医学研究基金会蛋白质序列数据库中的任何蛋白质都不密切相关，编码分泌蛋白质的血管性血友病因子cDNA序列的一部分也与Genbank基因组序列数据库中的任何序列都不同源。因此，血管性血友病因子中五种类型的重复结构域中有四种在其他已知蛋白质中没有同源物。四肽Arg-Gly-Asp-Ser出现在结构域C1的羧基末端，并可介导血管性血友病因子与活化血小板的GPIIb/IIIa复合物的结合。所有的结构域A1位于与静息血小板的GPIb结合的von Willebrand因子的50千道尔顿胰蛋白酶片段内[Fujimura，Y.，Titani，K.，Holland，LZ，Russell，S. R.，罗伯茨，J.R.，埃尔德，J.H.，Ruggeri，Z. M.，&齐默尔曼，T. S.（1986）J.Biol.Chem.261，381-385]。其余结构域（B、D和E）尚未与特定功能相关。血浆von Willebrand因子前体的氨基酸序列为His-Arg-Ser-Lys-Arg-Ser，成熟亚基是在Lys-Arg二肽后经蛋白水解裂解产生的。该序列类似于几种哺乳动物、病毒、真菌和酵母蛋白质前体的序列，其也在生物合成期间在配对碱性残基之后被蛋白水解加工。在本实验室测序的四个cDNA分离株中，有八个单核苷酸差异可能反映了血管性血友病因子基因序列的多态性。其中，七个是转换，一个是颠换。五个不影响翻译的蛋白质序列，但四个导致单个氨基酸取代。加上此前报道的...
Howard Hughes Medical Institute Laboratories, Departments of Medicine and Biochemistry, Washington University School of Medicine, St. Louis, Missouri 63110, and Department of Biochemistry, University of Washington, Seattle, Washington 98195 Received January 14, 1986; Revised Manuscript Received April 11, 1986 abstract: A human umbilical vein endothelial cell cDNA library in Xgtl 1 was screened with two previously described cDNA inserts for human von Willebrand factor. Among 16 positive isolates, two that hybridized with a probe corresponding to the amino terminus of von Willebrand factor were sequenced. Together, these four cDNA inserts span 6.5 kilobases of the von Willebrand factor mRNA sequence, completely specifying the 2050 amino acids of the subunit of mature, secreted von Willebrand factor and 24 residues of a precursor peptide. Approximately 77% of the sequence is contained in five types of repeated domains. Domain A consists of 193-220 amino acids and is present in three tandem copies between residues 497 and 1111. Domain B contains 25-35 amino acids and is present in three copies between residues 1533 and 1636. Domain C consists of 116-119 amino acids and is duplicated between residues 1637 and 1899. In contrast to the essentially contiguous repetition of domains AC, the two copies of domains D and E are each separated by 804 and 1383 amino acids, respectively. Domain D1 contains 289 amino acids between residues 79 and 367, while domain D2 consists of 270 amino acids between residues 1171 and 1440. Domain El consists of 46 amino acids between residues 25 and 70, and domain E2 consists of 46 amino acids between residues1453 and 1498. The triplicated A domains are notably poor inCys content, while the remaining domains are Cys-rich. The A domains appearto be homologous to a 225-residue segment of complement factor B. Otherwise, von Willebrand factor is not closely related to any protein in the National Biomedical Research Foundation Protein Sequence Database, nor is the portion of the von Willebrand factor cDNA sequence that encodes the secreted protein homologousto any sequence in the Genbank Genomic Sequence Data Bank. Thus, four of the five types of repeated domains in von Willebrand factor have no homologues among other known proteins. The tetrapeptide Arg-Gly-Asp-Ser occurs at the carboxy-terminal end of domain Cl and may mediate the binding of von Willebrand factor to the GPIIb/IIIa complex of activated platelets. All of domain Al lies within a 50-kilodalton tryptic fragment of von Willebrand factor that binds to GPIb of resting platelets [Fujimura, Y., Titani, K., Holland, LZ, Russell, S. R., Roberts, J. R., Elder, J. H., Ruggeri, Z. M., & Zimmerman, T. S.(1986) J. Biol. Chem. 261, 381-385]. The remaining domains (B, D, and E) have not been correlatedwith specific functions. The sequence of the von Willebrand factor precursor before the amino-terminal Ser of plasma von Willebrand factor is His-Arg-Ser-Lys-Arg-Ser. The mature subunit is generated by proteolytic cleavage after the Lys-Arg dipeptide. This sequence resembles that of several mammalian, viral, fungal, and yeast protein precursors that are also proteolytically processed after paired basic residues during biosynthesis. Among the four cDNA isolates sequenced by thislaboratory, there are eight single-nucleotide discrepancies that may reflect polymorphism in the von Willebrand factor gene sequence. Of these, seven are transitions, and one is a transversion. Five do not affect the translated protein sequence, but four result in single amino acid substitutions. Together with the previously reported …