Amino acid substitutions in conserved domains of factor VIII and related proteins: study of patients with mild and moderately severe hemophilia A.
Amino acid substitutions in conserved domains of factor VIII and related proteins: study of patients with mild and moderately severe hemophilia A.
复制标题
因子 VIII 和相关蛋白保守域的氨基酸取代：对轻度和中度重度血友病 A 患者的研究。
DOI：
10.1002/humu.1380010312
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发表时间：
1992
期刊：
影响因子：
3.9
通讯作者：
Gitschier,J
中科院分区：
文献类型：
--
作者：
Diamond,C;Kogan,S;Levinson,B;Gitschier,J
通过凝血因子VIII中氨基酸的取代导致血友病A的突变可能为这种大型且神秘的蛋白质的结构和功能提供重要线索。为了有效地发现错义突变，对患有轻度和中重度疾病的血友病患者进行了调查。在靶区域DNA扩增后，通过变性梯度凝胶电泳分析受影响个体的DNA样品的突变，所述靶区域包括除p53 B结构域之外的所有编码区。在检查的34名患者中有20名观察到错义突变，在5对患者中发现了相同的突变。所有突变均发生在重复A和C结构域。通过将因子VIII中的这些结构域与因子V、血浆铜蓝蛋白和小鼠乳脂球膜蛋白中的同源结构域进行比对，确定大多数突变改变强序列保守区域中的氨基酸。检测到另外三个突变，包括内含子中的点突变、终止密码子突变和沉默碱基变化。在该患者人群中发现的18种不同突变中，有10种是首次报道。© 1992 Wiley利斯公司
Mutations leading to hemophilia A by substitution of amino acids in coagulation factor VIII may provide important clues to the structure and function of this large and enigmatic protein. To efficiently find missense mutations, hemophiliacs with mild and moderately severe forms of the disease were surveyed. DNA samples from affected individuals were assayed for mutations by denaturing gradient gel electrophoresis following DNA amplification of target regions, which included all coding regions except for that of the dispensable B domain. Missense mutations were observed in 20 of the 34 patients examined, with identical mutations found in five pairs of patients. All mutations were found in the repetitive A and C domains. By aligning these domains in factor VIII with homologous domains in factor V, ceruloplasmin, and the mouse milk fat globule membrane protein, it was determined that most mutations change amino acids in areas of strong sequence conservation. Three additional mutations were detected, including a point mutation in an intron, a stop codon mutation, and a silent base change. Ten of the 18 different mutations discovered in this patient population are reported here for the first time. © 1992 Wiley‐Liss, Inc.