Expression and characterization of a human BMP-7 variant with improved biochemical properties
Expression and characterization of a human BMP-7 variant with improved biochemical properties
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DOI：
10.1016/j.pep.2007.09.016
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发表时间：
2008-02-01
影响因子：
1.6
通讯作者：
Amegadzie, Bernard
中科院分区：
文献类型：
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作者：
Swenckl-Underwood, Bethany;Mills, Juliane K.;Amegadzie, Bernard
骨形态发生蛋白-7 （BMP-7, OP-1）是一种分泌性生长因子，主要以其骨诱导特性而闻名，尽管它也被认为在哺乳动物肾脏发育中起作用。重组BMP-7外用治疗骨科损伤的临床疗效已得到证实。然而，用于全身给药的重组BMP-7的药物开发面临许多挑战。具体而言，重组成熟BMP-7蛋白在哺乳动物细胞中的表达水平非常低，该分子在中性pH下的溶解度较差，细胞内蛋白水解处理事件导致分泌的BMP-7具有多个氨基末端，形成异质蛋白混合物。利用结构信息，我们设计并产生了许多合理的BMP-7突变，这些突变提高了哺乳动物细胞中的表达水平和在中性pH下的溶解度，同时限制了成熟蛋白的氨基末端异质性。这些突变的引入并未影响BMP-7的体外生物活性。这种改进的BMP-7分子更适合于药物开发和需要全身给药的适应症的临床进展。(C) 2007爱思唯尔公司版权所有。
Bone morphogenetic protein-7 (BMP-7, OP-1) is a secreted growth factor that is predominantly known for its osteoinductive properties, though it has also been implicated as having a role in mammalian kidney development. Clinical efficacy of recombinant BMP-7 has been demonstrated in the treatment of orthopedic injuries through topical application. However, the pharmaceutical development of recombinant BMP-7 for systemic delivery has presented many challenges. Specifically, the expression level of recombinant mature BMP-7 protein in mammalian cells is very low, the molecule has poor solubility at neutral pH, and intracellular proteolytic processing events result in a secreted BMP-7 having multiple amino-termini, creating a heterogeneous mixture of proteins. Utilizing structural information, we have designed and generated a number of rational BMP-7 mutations that improved both expression levels in mammalian cells and solubility at neutral pH, while limiting the amino-terminal heterogeneity of the mature protein. Introduction of these mutations did not compromise BMP-7 in vitro bioactivity. This improved BMP-7 molecule is better suited for pharmaceutical development and clinical advancement for indications where systemic delivery may be required. (C) 2007 Elsevier Inc. All rights reserved.