High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum.
High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum.
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血小板源性内皮细胞生长因子的高产纯化：结构表征和特异性抗血清的建立。
DOI：
10.1021/bi00430a042
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发表时间：
1989
期刊：
影响因子：
2.9
通讯作者：
C. Heldin
中科院分区：
文献类型：
--
作者：
K. Miyazono;C. Heldin
血小板源性内皮细胞生长因子 (PD-ECGF) 是一种 45 kDa 的蛋白质，可刺激内皮细胞生长 [Miyazono, K., et al. 2017]。 (1987) J.Biol。化学。 262, 4098-4103]。在这里，我们描述了一种从人血小板裂解物中高产率纯化大量 PD-ECGF 的方法（总回收率 14%）。该纯化方法涉及五个步骤，使用高效液相色谱级羟基磷灰石和疏水色谱作为最后两个步骤。纯化的材料含有表观分子量值为46,000和44,000的两种主要成分。这些成分在高分辨率反相色谱中共洗脱，并在用葡萄球菌 V8 蛋白酶处理后发现相似的肽图，表明 44-kDa 形式与 46-kDa 分子相关。天然 PD-ECGF 的部分胰蛋白酶消化显示该分子含有 37-39 kDa 的胰蛋白酶抗性结构域。针对纯化的材料产生了兔抗血清，并发现在免疫印迹中特异性识别 PD-ECGF。当添加到细胞培养基中时，抗血清的免疫球蛋白部分中和了纯化的 PD-ECGF 的活性。此外，它完全中和了血小板裂解物的内皮细胞促有丝分裂活性，表明PD-ECGF是血小板裂解物中针对该细胞类型的唯一有丝分裂原。
Platelet-derived endothelial cell growth factor (PD-ECGF) is a 45-kDa protein that stimulates the growth of endothelial cells [Miyazono, K., et al. (1987) J. Biol. Chem. 262, 4098-4103]. Here, we describe a method to purify large quantities of PD-ECGF from human platelet lysate at a high yield (14% overall recovery). The purification method involves five steps, using high-performance liquid chromatography grade hydroxylapatite and hydrophobic chromatographies as the two final steps. The purified material contained two major components of apparent molecular weight values of 46,000 and 44,000. These components coeluted in a high-resolving reversed-phase chromatography and were found to give similar peptide maps after treatments with staphylococcal V8 protease, suggesting that the 44-kDa form is related to the 46-kDa molecule. Partial tryptic digestion of native PD-ECGF revealed that the molecule contains a trypsin-resistant domain of 37-39 kDa. A rabbit antiserum was produced against the purified material and was found to specifically recognize PD-ECGF in immunoblotting. When added to the cell culture medium, an immunoglobulin fraction of the antiserum neutralized the activity of purified PD-ECGF. Furthermore, it completely neutralized the endothelial cell mitogenic activity of platelet lysate, indicating that PD-ECGF is the only mitogen in platelet lysate for this cell type.
影响因子：
56.9
作者：
HEIMARK, RL;TWARDZIK, DR;SCHWARTZ, SM
通讯作者：
SCHWARTZ, SM
肝素结合内皮细胞生长因子的纯化和表征。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lobb,R;Sasse,J;Sullivan,R;Shing,Y;D'Amore,P;Jacobs,J;Klagsbrun,M
通讯作者：
Klagsbrun,M
DOI：
10.1016/0006-291x(86)90305-0
发表时间：
1986-07-16
影响因子：
3.1
作者：
BAIRD, A;DURKIN, T
通讯作者：
DURKIN, T