Vascular regulation by kringles of HGF and molecular cloning of a novel kringle-containing protein

含有Kringle结构域的各种蛋白质在组织再生、凝血/纤溶、神经发育和血管调节中发挥着不同的作用。一个新的Kringle分子的鉴定可能导致对血管调节的进一步了解，包括血管生成和抗血管生成。在本研究中，为了研究Kringle结构域发挥血管调节功能的调节机制，我们分析了NK4的血管抑制作用，NK4以前被认为是HGF拮抗剂。同样，我们成功地克隆了一种新的含kringle的分子Kremen1)NK4作为血管生成抑制剂：NK4是一种含四kringle的HGF拮抗剂，可抑制培养的血管内皮细胞的增殖和迁移。同样，NK4在体内抑制了CAM和角膜中的血管生成。结合nk4作为肝细胞生长因子拮抗剂的生物学作用，nk4治疗转移性c…的潜力更多的舞者接受了测试。给小鼠注射NK4可以有效地抑制肺癌、乳腺癌和胰腺癌等癌症的转移、肿瘤血管生成和肿瘤生长。NK4将成为一种有效的抗癌药物，针对癌症的恶性行为。2)利用一种新建立的Kringle-Sage蛋白克隆策略，我们克隆了Kreman的cDNA.克雷曼蛋白由473个氨基酸组成，为I型跨膜蛋白。Kreman在其胞外区有独特的结构域、Kringle结构域和Cub结构域，而在其胞内区没有特定的保守功能基序。克雷曼基因在多种组织中表达，在肌肉和神经分化过程中表达上调。Kreman的Kringle结构域可能在血管调节和肌肉和神经细胞的分化中发挥一些作用，但其作用仍有待进一步研究。Kringle-SAGE是一种新建立的全面克隆Kringle蛋白的方法，在Kringle蛋白的分子克隆中具有强大的功能。较少

Various proteins containing kringle domains play diverse roles in tissue regeneration, blood coagulation/fibrinolysis, neural development, and blood vessel regulation. Identification of a novel kringle-containing molecule is likely to lead to further understanding of blood vessel regulation, including angiogenesis and anti-angiogenesis. In the present study, to study regulatory mechanisms by which kringle domains exhibit angio-regulatory functions, we analyzed angiostatic action of NK4, previously identified as HGF-antagonist. Likewise, we succeeded in molecular cloning of a novel kringle-containg molecule, Kremen.1) NK4 as angiogenesis inhibitor : NK4, a four kringles-cpontaining HGF-antagonist, inhibited proliferation and migration of vascular endothelial cells in culture. Likewise, NK4 inhibited angiogenesis in CAM and cornea assays in vivo. Together with a previously characterized biological action of NK4 as HGF-antagonist, therapeutic potential of NK4 for treatment of metastatic c … More ancers were tested. NK4 administered into mice potently suppressed metastasis, tumor angiogenesis, and tumor growth of cancers, including lung, breast, and pancreatic cancers. NK4 will become a potent anti-cancer agent, targeting malignant behavior of cancer.2) Using a newly established cloning strategy for kringle-containg proteins, 'Kringle-SAGE', we cloned cDNA for Kremen. Kremen is composed of 473 amino acids and is type-I transmembrane protein. Kremen contains unique domains, kringle domain and CUB domain in its extracellular region, while it has no specific conserved functional motif in its intracellular region. Kremen mRNA is expressed in diverse tissues, while it is up-regulated during muscular and neural differentiation. Kringle domain of Although Kremen may have some roles in vascular regulation and differentiation of muscular and neural cells, its roles remain to be further addressed. A newly established method for comprehensive cloning of kringle-proteins, Kringle-SAGE is powerful in molecular cloning of kringle proteins. Less