BMP-1-like protease effects on growth factors & hormones

BMP-1 样蛋白酶对生长因子的影响

• 批准号: 6931027
• 负责人: DANIEL S GREENSPAN
• 金额: $ 25.99万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931027
• 关键词: bone morphogenetic proteins; chemical cleavage; endopeptidases; gene targeting; genetic manipulation; glycoproteins; growth factor; hormones; inhibitor /antagonist; laboratory mouse; myostatin; prolactin; protein protein interaction; protein sequence; protein structure function; proteolysis; somatomammotropin; somatotropin; transforming growth factors

DESCRIPTION (provided by applicant): The 4 mammalian BMP-l-like proteases (BLPs) play central roles in forming the ECM through biosynthetic processing of precursors, and enhance BMP signaling by cleaving the extracellular antagonist Chordin. Here we show BLPs to cleave prodomain sequences of the TGF-b-like factor gdf-8 (myostatin). Secreted gdf-8 is normally noncovalently bound to its prodomain in a latent complex. Cleavage of prodomain sequences by BLPs is shown to activate gdf-8. Golf-8 inhibits muscle growth, and mutation of the prodomain BLP cleavage site is shown to make the prodomain a powerful stimulator of muscle growth when administered to adult mice. Thus, cleavage at this site is biologically important. We propose showing which BLP may be responsible for activating gdf-8 in vivo. Gdf-11 (BMP-11) and gdf-8 form a subfamily of TGF-b-like molecules based on sequence homologies in their mature regions. We have preliminary evidence that the gdf-11 propeptide is cleaved by BLPs. We propose showing that mature secreted gdf-11 noncovalently binds its prodomain, that BLPs cleave the prodomain to release and activate mature gdf-11, and that this occurs in vivo. Gdf-11 is an inhibitor of neural tissue formation. Therefore, characterizing the mechanisms for in vivo activation of gdfs 8 and 11 has important implications for how such activation might be blocked, and thus how muscle and nerve regeneration may be enhanced in musculo- and neuro-degenerative diseases. We also propose determining which BI-Ps may cleave extracellular BMP antagonists Chordin-like 1 and 2, which show much overall homology with Chordin and which show indications of being regulated via proteolysis. In addition, we present evidence that BLPs may be involved in processing members of the prolactin, growth hormone/placental lactogen family of hormones to produce specific 16 kDa product. We propose determining which of these hormones are cleaved by which BLPs, at which sites, and whether such cleavages occur in vivo. Previous reports indicate that 16 kDa products of these hormones to biologically important. Thus, determining how these fragments are formed is of potential importance in regulating human endocrine functions. Conditional knockout mice for BLP genes will be created, for in vivo portions of the projects described above, and for future studies of the tissue-specific roles of BLPs in development and homeostasis. Finally, we present evidence for and propose experiments to further determine in vivo roles for the precursor forms and prodomains of BLPs in development and homeostasis.

描述（由申请人提供）：4种哺乳动物BMP-1样蛋白酶（BLP）在通过前体的生物合成加工形成ECM中发挥核心作用，并通过裂解细胞外拮抗剂脊索蛋白来增强BMP信号传导。在这里，我们展示了 BLP 可以切割 TGF-b 样因子 gdf-8（肌生长抑制素）的前结构域序列。分泌的 gdf-8 通常在潜在复合物中与其前结构域非共价结合。 BLP 对前结构域序列的切割可激活 gdf-8。 Golf-8 抑制肌肉生长，并且当给予成年小鼠时，前结构域 BLP 裂解位点的突变显示出使前结构域成为肌肉生长的强大刺激剂。因此，该位点的切割具有重要的生物学意义。我们建议展示哪个 BLP 可能负责体内激活 gdf-8。 Gdf-11 (BMP-11) 和 gdf-8 基于其成熟区域的序列同源性形成 TGF-b 样分子的亚家族。我们有初步证据表明 gdf-11 前肽被 BLP 切割。我们建议证明成熟的分泌型 gdf-11 非共价结合其前结构域，BLP ​​裂解前结构域以释放并激活成熟的 gdf-11，并且这在体内发生。 Gdf-11 是神经组织形成的抑制剂。因此，表征 gdfs 8 和 11 的体内激活机制对于如何阻止这种激活以及如何在肌肉和神经退行性疾病中增强肌肉和神经再生具有重要意义。我们还建议确定哪些 BI-P 可以裂解细胞外 BMP 拮抗剂 Chordin 样 1 和 2，它们显示出与 Chordin 的总体同源性，并且显示出通过蛋白水解进行调节的迹象。此外，我们提供的证据表明，BLP ​​可能参与催乳素、生长激素/胎盘催乳激素家族成员的加工，以产生特定的 16 kDa 产物。我们建议确定这些激素中的哪些被哪些 BLP 在哪些位点裂解，以及这种裂解是否在体内发生。先前的报告表明这些激素的 16 kDa 产物具有重要的生物学意义。因此，确定这些片段是如何形成的对于调节人类内分泌功能具有潜在的重要性。将创建 BLP 基因的条件性敲除小鼠，用于上述项目的体内部分，以及未来研究 BLP 在发育和稳态中的组织特异性作用。 最后，我们提供证据并提出实验，以进一步确定 BLP 的前体形式和前结构域在发育和稳态中的体内作用。