A LARGE REGION (ALMOST-EQUAL-TO-95 KDA) OF HUMAN FACTOR-VIII IS DISPENSABLE FOR INVITRO PROCOAGULANT ACTIVITY
A LARGE REGION (ALMOST-EQUAL-TO-95 KDA) OF HUMAN FACTOR-VIII IS DISPENSABLE FOR INVITRO PROCOAGULANT ACTIVITY
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DOI：
10.1073/pnas.83.16.5939
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发表时间：
1986-08-01
影响因子：
11.1
通讯作者：
KAUFMAN, RJ
中科院分区：
文献类型：
--
作者：
TOOLE, JJ;PITTMAN, DD;KAUFMAN, RJ
因子 VIII（抗血友病因子）是一种参与凝血级联的高分子量血浆糖蛋白。最近对编码人因子VIII的cDNA进行克隆和序列分析揭示了该蛋白明显的结构域结构，可以表示为A1-A2-B-A3-C1-C2。我们现在报告编码整个 B 结构域以及部分 A2 和 A3 结构域的猪外显子的 DNA 序列分析。我们发现，与猪因子 VIII 分子其他区域的有限序列相比，B 区域的猪与人氨基酸序列差异异常高。除了序列差异之外，猪 B 结构域中还存在大量缺口，总计超过 200 个氨基酸。使用重组DNA技术从全长人因子VIII cDNA中去除编码B结构域的大片段DNA。尽管删除了高达 38% 的因子 VIII 分子，但这些构建体在引入哺乳动物细胞时仍指导生物活性因子 VIII 的合成。
Factor VIII (antihemophilic factor) is a high molecular weight plasma glycoprotein that participates in the blood clotting cascade. The recent cloning and sequence analysis of the cDNA encoding human factor VIII revealed an obvious domain structure for the protein, which can be represented as A1-A2-B-A3-C1-C2. We now report the DNA sequence analysis of porcine exons encoding the entire B domain and part of the A2 and A3 domains. We found an unusually high degree of porcine-human amino acid sequence divergence in the B region compared with the limited sequence available for other regions of the porcine factor VIII molecule. In addition to sequence divergence, there are numerous gaps in the porcine B domain totalling over 200 amino acids. Recombinant DNA techniques were used to effect the removal of large segments of DNA encoding the B domain from the full length human factor VIII cDNA. These constructs directed the synthesis of biologically active factor VIII when introduced into mammalian cells despite the deletion of up to 38% of the factor VIII molecule.