Isolation of an amino terminal extended form of basic fibroblast growth factor.
Isolation of an amino terminal extended form of basic fibroblast growth factor.
复制标题
碱性成纤维细胞生长因子的氨基末端延伸形式的分离。
DOI：
10.1016/s0006-291x(86)80536-8
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发表时间：
1986
影响因子：
3.1
通讯作者：
Guillemin,R
中科院分区：
文献类型：
--
作者：
Ueno,N;Baird,A;Esch,F;Ling,N;Guillemin,R
在酶抑制剂（2 mM PMSF、2 mM 连四硫酸钠、15 μM 胃酶抑素 A 和 1 mM EDTA）存在下提取牛垂体，分离出两种不同形式的碱性成纤维细胞生长因子。两种分子的部分表征表明其中一种形式与我们实验室已报道的基本 FGF(1-146) 相同。通过SDS-PAGE估计第二种形式的分子量为17,000道尔顿，略大于碱性FGF(1-146)。氨基酸分析表明存在比碱性 FGF(1-146) 多的 8 个新残基，这是造成分子量差异的原因。该分子的气相测序表明它具有封闭的氨基末端。此外，这种较高分子量形式的碱性 FGF 不显示与碱性 FGF(1-146) 氨基末端特异性抗体的免疫反应性，但与针对碱性 FGF(1-146) 中部片段产生的抗体发生交叉反应，表明该分子是氨基末端延伸的。与碱性 FGF(1-146) 一样，该分子是血管内皮细胞的有效有丝分裂因子。总而言之，这些结果证明存在碱性 FGF 的前体形式，其在氨基末端延伸 8 个残基，其中第一个残基被封闭。
Extraction of bovine pituitaries in the presence of enzyme inhibitors (2 mM PMSF, 2 mM sodium tetrathionate, 15 μM pepstatin A, and 1 mM EDTA) resulted in the isolation of two distinct forms of basic fibroblast growth factor. Partial characterization of both molecules showed one form to be identical to basic FGF(1-146) which has already been reported by our laboratory. The second form was estimated by SDS-PAGE to have a molecular weight of 17,000 Daltons which is slightly larger than that of basic FGF(1-146). Amino acid analysis shows the presence of 8 new residues more than basic FGF(1-146) which accounts for the difference in molecular weight. Gas-phase sequencing of this molecule indicated that it bears a blocked amino terminus. Furthermore, this higher molecular weight form of basic FGF did not show immunoreactivity with antibodies specific for the amino terminus of basic FGF(1-146) but cross reacted with antibodies generated against midportion fragments of basic FGF(1-146), indicating that the molecule is amino terminally extended. Like basic FGF(1-146), the molecule is a potent mitogenic factor for vascular endothelial cells. Taken together these results demonstrate the existence of a precursor form of basic FGF which is extended by 8 residues at the amino terminus with the first residue being blocked.