Expression and purification of recombinant protein related to DAN and cerberus (PRDC)

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• 作者：Chandramohan Kattamuri ; David M. Luedeke ; Thomas B. Thompson ; ^{tom.thompson@ucmail.uc.edu} ; ^{tom.thompson@uc.edu}
• 关键词：Oxidative refolding ; BMP antagonist ; TGF尾 family
• 刊名：Protein Expression and Purification
• 出版年：2012
• 期刊代码：184_10465928
• 类别：bio
• 出版时间：April, 2012
• 卷：82
• 期：2
• 页码：389-395
• 文件大小：559 K

摘要

Bone morphogenetic proteins (BMPs) are secreted protein ligands that control numerous biological processes, such as cell differentiation and cell proliferation. Ligands are regulated by a large number of structurally diverse extracellular antagonists. PRDC or protein related to DAN and cerberus is a BMP antagonist of the DAN family, which is defined by a conserved pattern of cysteine residues that form a ring structure. Here we present the expression and purification of recombinant mouse PRDC (mPRDC) from bacterial (Escherichia coli) inclusion bodies through oxidative refolding. Functional mPRDC was isolated from a nonfunctional component through reverse phase chromatography and shown to inhibit BMP2 and BMP4 in a cell-based luciferase reporter assay. Recombinant mPRDC also bound directly to BMP2, BMP4 and BMP7, but not activin A. Furthermore, circular dichroism indicated that mPRDC is folded and contains a higher than anticipated helical content for a DAN family member protein.