A fusion protein of hepatocyte growth factor for immobilization to collagen
A fusion protein of hepatocyte growth factor for immobilization to collagen
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DOI：
10.1016/j.biomaterials.2006.12.022
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发表时间：
2007-04-01
期刊：
影响因子：
14
通讯作者：
Ito, Yoshihiro
中科院分区：
文献类型：
--
作者：
Kitajima, Takashi;Terai, Hiroshi;Ito, Yoshihiro
我们在这里描述的融合蛋白组成的肝细胞生长因子（HGF;血管生成因子）和胶原结合域（CBD）的纤维连接蛋白（FN）多肽。由杆状病毒表达系统产生的这种融合蛋白（CBD-HGF）在0.4 - 6.4 μ g/ml的范围内表现出比天然HGF强得多的胶原结合活性。其在最低浓度下的结合超过在最高浓度下的HGF的结合。此外，与胶原蛋白结合的CBD-HGF比添加到培养基中的HGF至少多4天，在更大程度上促进了内皮细胞（EC）的生长; 10天后细胞数量增加约5倍。这些发现表明，融合的CBD部分不仅有助于将HGF粘附在胶原蛋白上，而且有助于稳定融合分子，从而延长活性。通过皮下植入含有结合的CBD-HGF的胶原海绵来检查CBD-HGF在动物组织中的血管生成活性。与没有样品的对照海绵相比，7天后海绵中的血管形成是4 - 6倍。植入天然HGF的海绵与对照组相比没有显着差异。这些结果表明CBD-HGF适合于EC的体外培养，并且该融合蛋白可用于将HGF活性赋予用于组织工程的生物材料。(c)2007爱思唯尔有限公司保留所有权利。
We describe here a fusion protein consisting of hepatocyte growth factor (HGF; an angiogenic factor) and a collagen-binding domain (CBD) polypeptide of fibronectin (FN). This fusion protein (CBD-HGF), produced by a baculovirus expression system, exhibited much stronger collagen binding activity than native HGF in the range of 0.4-6.4 mu g/ml. Its binding at the lowest concentration exceeded that of HGF at the highest concentration. In addition, the collagen-bound CBD-HGF promoted growth of endothelial cells (ECs) to a greater degree at least 4 days longer than HGF added to the culture medium; about 5-fold greater increase in cell number after 10 days. These findings suggest that the fused CBD moiety not only helped immobilize HGF on collagen but also helped stabilize the fusion molecule, resulting in prolonged activity. The angiogenic activity of CBD-HGF in animal tissues was examined by subcutaneously implanting collagen sponges containing bound CBD-HGF. Blood vessel formation in the sponges after 7 days was 4-6-fold extensive as compared to the control sponges without sample. Implanted sponges with native HGF did not show significant difference from control. These results indicate that CBD-HGF is suitable for in vitro culture of ECs, and that this fusion protein can be used to confer HGF activity on biornaterials for use in tissue engineering. (c) 2007 Elsevier Ltd. All rights reserved.