BIOLOGICAL ACTIVITY AND STRUCTURAL ANALYSIS OF KGF, HGF AND SECRETED FRIZZLED REL

KGF、HGF和分泌型卷曲相关蛋白的生物活性和结构分析

• 批准号: 6289320
• 负责人: JEFFREY RUBIN
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6289320
• 关键词: fibroblast growth factor; fluorescent in situ hybridization; gene expression; gene mutation; hepatocyte growth factor; nuclear magnetic resonance spectroscopy; protein structure function; tissue /cell culture

During the past year, we have continued to focus our research efforts on the study of a secreted, Frizzled-related protein (sFRP-1) that is thought to have an important role in embryonic development and perhaps neoplasia. We developed an abundant source of recombinant sFRP-1 to demonstrate that it can bind directly to Wnt protein and exert a biphasic effect on Wnt activity. Several collaborative studies with recombinant sFRP-1 are broadening our understanding of its biological activity. In a rat embryonic kidney organ culture, sFRP-1 can block tubulogenesis presumably by inhibiting Wnt-4 activity. Preliminary results suggest that sFRP-1 alters the behavior of stem cells from the rat central nervous system and microvascular endothelial cells from rat kidney. We also have been investigating sfrp-1 gene structure and characterizing its promoter activity to better understand how its expression is regulated.Substantial progress has been made in an ongoing collaboration concerning the structure-function analysis of HGF/SF. NMR solution studies involving the amino-terminal domain and heparin or heparin analogs have yielded new information about the primary heparin-binding site of HGF/SF. Additional experiments are in progress to confirm the identity of the amino acid residues responsible for heparin-binding, and to determine the consequences of their replacement with residues that will not mediate heparin interactions. Collaborative studies have yielded new information about the activity of keratinocyte growth factor (KGF or FGF-7). In particular, KGF was shown to stimulate the growth of spiral arteries in primate uterus, which could be important for the establishment of fetal-maternal circulation. Other ongoing investigations concern the role of KGF/KGFR signaling in thymus development and T cell maturation, and the mechanisms responsible for KGF?s ability to ameliorate the impact of graft versus host disease associated with bone marrow transplantation. - cell proliferation, growth factor, HGF/SF, protein function, receptor, Wnt signaling, Frizzled, KGF,

在过去的一年中，我们继续把我们的研究工作集中在一种分泌的卷曲相关蛋白（sFRP-1）的研究上，这种蛋白被认为在胚胎发育和肿瘤形成中起重要作用。我们开发了丰富的重组sFRP-1的来源，以证明它可以直接结合Wnt蛋白，并对Wnt活性产生双相作用。一些与重组sFRP-1的合作研究正在扩大我们对其生物活性的理解。在大鼠胚胎肾器官培养中，sFRP-1可能通过抑制Wnt-4活性来阻断肾小管形成。初步结果表明sFRP-1改变了大鼠中枢神经系统干细胞和大鼠肾脏微血管内皮细胞的行为。我们也一直在研究sfrp-1基因的结构，并表征其启动子活性，以更好地了解其表达是如何调节的。涉及氨基末端结构域和肝素或肝素类似物的NMR溶液研究已经产生了关于HGF/SF的主要肝素结合位点的新信息。正在进行其他实验，以确认负责肝素结合的氨基酸残基的身份，并确定将其替换为不会介导肝素相互作用的残基的后果。合作研究已经产生了关于角质细胞生长因子（KGF或FGF-7）活性的新信息。特别是，KGF被证明可以刺激灵长类动物子宫中螺旋动脉的生长，这对于建立胎儿-母体循环可能是重要的。其他正在进行的调查关注的作用，KGF/KGFR信号在胸腺发育和T细胞成熟，和机制负责KGF？改善骨髓移植相关的移植物抗宿主病的影响的能力。- 细胞增殖，生长因子，HGF/SF，蛋白质功能，受体，Wnt信号传导，卷曲，KGF，