Biological Activity and Structural Analysis of Secreted Frizzled-Related Protein

分泌型卷曲相关蛋白的生物活性及结构分析

• 批准号: 6433205
• 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/6433205
• 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. Our recent work with other recombinant sFRP family members indicates differences in their biological activities and functions. 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. 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). 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.

在过去的一年中，我们继续将研究重点放在研究一种分泌的卷曲相关蛋白（sFRP-1）上，该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来源，以证明它可以直接结合到Wnt蛋白上，并对Wnt活性发挥双相作用。一些与重组sFRP-1的合作研究正在拓宽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活性来阻断小管形成。我们最近对其他重组sFRP家族成员的研究表明，它们的生物活性和功能存在差异。我们也一直在研究条带-1基因结构和表征其启动子活性，以更好地了解其表达是如何被调节的。关于HGF/SF结构功能分析的合作已经取得了实质性进展。实验正在进行中，以确认负责肝素结合的氨基酸残基的身份，并确定它们被不介导肝素相互作用的残基取代的后果。合作研究已经获得了关于角质细胞生长因子（KGF或FGF-7）活性的新信息。研究涉及KGF/KGFR信号在胸腺发育和T细胞成熟中的作用，以及KGF改善骨髓移植相关移植物抗宿主病影响的能力的机制。