Expression, purification and osteogenic bioactivity of recombinant human BMP-4,-9,-10,-11 and-14
Expression, purification and osteogenic bioactivity of recombinant human BMP-4,-9,-10,-11 and-14
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DOI：
10.1016/j.pep.2008.09.014
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发表时间：
2009-02-01
影响因子：
1.6
通讯作者：
Casal, M.
中科院分区：
文献类型：
--
作者：
Bessa, P. C.;Cerqueira, M. T.;Casal, M.
骨形态发生蛋白（BMPs）是来自TGF-β超家族的细胞因子，在胚胎发育期间以及在成人体内诱导骨和软骨组织分化中具有重要作用。本文报道了利用大肠杆菌pET-25 b载体表达重组人骨形态发生蛋白（BMPA）、骨形态发生蛋白（BMP-9）、骨形态发生蛋白（BMP-10）、骨形态发生蛋白（BMP-11）（或生长分化因子（GDF-11））和骨形态发生蛋白（BMP-14）（GDF-5）。BMP过表达，通过亲和组氨酸标签色谱纯化，并显示诱导C2 C12细胞和人脂肪干细胞中骨分化的早期标志物（例如，smad-1、smad-5、runx 2/cbfa 1、dlx 5、osterix、骨桥蛋白、骨唾液酸蛋白和碱性磷酸酶）的表达。所描述的方法是一种很有前途的方法，用于生产大量的不同的重组骨形成蛋白，显示出潜在的新的生物医学应用。(C)2008年爱思唯尔公司All rights reserved.
Bone morphogenetic proteins (BMPs) are cytokines from the TGF-beta superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMPA, BMP-9, BMP-10, BMP-11 (or growth differentiation factor-11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications. (C) 2008 Elsevier Inc. All rights reserved.