AMINO-ACID-SEQUENCE OF THE B-SUBUNIT OF HUMAN FACTOR-XIII, A PROTEIN COMPOSED OF 10 REPETITIVE SEGMENTS
AMINO-ACID-SEQUENCE OF THE B-SUBUNIT OF HUMAN FACTOR-XIII, A PROTEIN COMPOSED OF 10 REPETITIVE SEGMENTS
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DOI：
10.1021/bi00364a027
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发表时间：
1986-08-12
期刊：
影响因子：
2.9
通讯作者：
DAVIE, EW
中科院分区：
文献类型：
--
作者：
ICHINOSE, A;MCMULLEN, BA;DAVIE, EW
因子XIII是一种血浆蛋白，参与血液凝固的最后阶段。通过cDNA克隆和氨基酸序列分析的组合，确定了人因子XIII的B亚基的完整氨基酸序列。用亲和纯化的抗人因子XIII的B亚基的抗体筛选从人肝mRNA制备的cDNA文库。从2 × 10 - 6细胞中分离出9个阳性克隆。106噬菌体和噬斑纯化。对最大的cDNA插入片段进行测序，结果显示含有2180个碱基对，编码前导序列的一部分（19个氨基酸）、成熟蛋白（641个氨基酸）、终止密码子（TGA）、3“非编码区（187个核苷酸）和poly（A）尾。当人因子XIII的B亚基用溴化氰消化时，通过凝胶过滤和反相高效液相色谱法分离出9种肽。用自动测序仪对这些肽进行氨基酸序列分析，鉴定了299个氨基酸残基。这些氨基酸序列与从cDNA预测的氨基酸序列完全一致。因子XIII的B亚基含有10个重复的同源片段，每个片段由约60个氨基酸和4个半胱氨酸残基组成。这些重复片段中的每一个都是存在于人β 2-糖蛋白I、补体因子B和触珠蛋白α 1链中的重复序列家族的成员。在因子XIII的B亚基中也发现了三个潜在的天冬酰胺连接的碳水化合物附着位点。
Factor XIII is a plasma protein that participates in the final stages of blood coagulation. The complete amino acid sequence of the b subunit of human factor XIII was determined by a combination of cDNA cloning and amino acid sequence analysis. A .lambda.gt11 cDNA library prepared from human liver mRNA was screened with an affinity-purified antibody against the b subunit of human factor XIII. Nine positive clones were isolated from 2 .times. 106 phage and plaque-purified. The largest cDNA insert was sequenced and shown to contain 2180 base pairs coding for a portion of the leader sequence (19 amino acids), the mature protein (641 amino acids), a stop codon (TGA), a 3'' noncoding region (187 nucleotides), and a poly(A) tail. When the b subunit of human factor XIII was digested with cyanogen bromide, nine peptides were isolated by gel filtration and reverse-phase high-performance liquid chromatography. Amino acid sequence analyses of these peptides were performed with an automated sequenator, and 299 amino acid residues were identified. These amino acid sequences were in complete agreement with the amino acid sequence predicted from the cDNA. The b subunit of factor XIII contained 10 repetitive homologous segments, each composed of about 60 amino acids and 4 half-cystine residues. Each of these repeated segments is a member of a family of repeats present in human .beta.2-glycoprotein I, complement factor B, and haptoglobin .alpha.1 chain. Three potential Asn-linked carbohydrate attachment sites were also identified in the b subunit of factor XIII.