Institute for Biochemistry, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
J Biol Chem. 2013 Mar 8;288(10):7193-203. doi: 10.1074/jbc.M112.433029. Epub 2013 Jan 22.
Factor VII-activating protease (FSAP) is a circulating protease involved in the pathogenesis of atherosclerosis, calcification, and fibrotic processes. To understand how FSAP controls the balance of local growth factors, we have investigated its effect on the regulation of bone morphogenetic proteins (BMPs). BMP-2 is produced as a large pro-form and secreted as a mature heparin-binding growth factor after intracellular processing by pro-protein convertases (PCs). In this study, we discovered that FSAP enhances the biological activity of mature BMP-2 as well as its pro-form, as shown by osteogenic differentiation of C2C12 myoblasts. These findings were complemented by knockdown of FSAP in hepatocytes, which revealed BMP-2 processing by endogenous FSAP. N-terminal sequencing indicated that pro-BMP-2 was cleaved by FSAP at the canonical PC cleavage site, giving rise to mature BMP-2 (Arg(282)↓Gln(283)), as well as in the N-terminal heparin binding region of mature BMP-2, generating a truncated mature BMP-2 peptide (Arg(289)↓Lys(290)). Similarly, mature BMP-2 was also cleaved to a truncated peptide within its N-terminal region (Arg(289)↓Lys(290)). Plasmin exhibited a similar activity, but it was weaker compared with FSAP. Thrombin, Factor VIIa, Factor Xa, and activated protein C were not effective. These results were further supported by the observation that the mutation of the heparin binding region of BMP-2 inhibited the processing by FSAP but not by PC. Thus, the proteolysis and activation of pro-BMP-2 and mature BMP-2 by FSAP can regulate cell differentiation and calcification in vasculature and may explain why polymorphisms in the gene encoding for FSAP are related to vascular diseases.
因子 VII 激活蛋白酶(FSAP)是一种循环蛋白酶,参与动脉粥样硬化、钙化和纤维化过程的发病机制。为了了解 FSAP 如何控制局部生长因子的平衡,我们研究了其对骨形态发生蛋白(BMPs)调节的影响。BMP-2 以大前体形式产生,并在蛋白原转化酶(PCs)的细胞内加工后作为成熟的肝素结合生长因子分泌。在这项研究中,我们发现 FSAP 增强了成熟 BMP-2 及其前体的生物学活性,正如 C2C12 成肌细胞的成骨分化所表明的那样。这些发现得到了肝细胞中 FSAP 敲低的补充,这揭示了内源性 FSAP 对 BMP-2 的加工。N 端测序表明,FSAP 在经典的 PC 切割位点切割前体 BMP-2,产生成熟的 BMP-2(Arg(282)↓Gln(283)),以及成熟 BMP-2 的 N 端肝素结合区,产生截短的成熟 BMP-2 肽(Arg(289)↓Lys(290))。同样,成熟的 BMP-2 也在其 N 端区域被切割成截短的肽(Arg(289)↓Lys(290))。纤溶酶也表现出类似的活性,但比 FSAP 弱。凝血酶、因子 VIIa、因子 Xa 和激活蛋白 C 无效。这些结果进一步得到了观察结果的支持,即 BMP-2 的肝素结合区突变抑制了 FSAP 的加工,但不抑制 PC 的加工。因此,FSAP 对前体 BMP-2 和成熟 BMP-2 的蛋白水解和激活可以调节血管中的细胞分化和钙化,并可以解释为什么编码 FSAP 的基因的多态性与血管疾病有关。