FGF 1 SECRETION AND THE PATHOLOGY OF ANGIOGENESIS

FGF 1 分泌和血管生成的病理学

• 批准号: 6030539
• 负责人: THOMAS MACIAG
• 金额: $ 20.28万
• 依托单位: MAINEHEALTH
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6030539
• 关键词: SDS polyacrylamide gel electrophoresis; angiogenesis; antibody formation; binding proteins; dimer; endopeptidases; exocytosis; fibroblast growth factor; fluorescence spectrometry; gel mobility shift assay; growth factor receptors; immunoprecipitation; intracellular transport; laboratory rabbit; posttranslational modifications; protein purification; protein sequence; protein signal sequence; protein structure function; protein transport; secretory protein; synaptotagmin; temperature; western blottings

DESCRIPTION (Adapted from the Investigator's Abstract): Heparin-binding fibroblast growth factor (FGF)-1 is a potent modifier of angiogenic and neurotrophic phenomena in vivo which signals these responses by including tyrosine phosphorylation of a family of high affinity FGF receptors (R) on the surface of target cells. Because FGF-1 does not contain a classical signal peptide sequence to direct its secretion through the endoplasmic reticulum-Golgi apparatus, yet requires FGFR signaling for function, it is important to understand the mechanism by which FGF-1 is secreted. They have previously shown that FGF-1 is released as a biologically inactive and poor heparin-binding protein in response to temperature stress in vitro. Additional studies from this laboratory have shown that FGF-1 (i) can be activated by reduced glutathione, (ii) is released as a cys30-cys30 FGF-1 homodimer, (iii) is able to associate with phosphatidylserine with high affinity, and (iv) is complexed to a cytosolic p45 fragment of p65 synaptotagmin (stg)-1 in both temperature-conditioned medium and in ovine brain. Interestingly, p65 stg-1 is a transmembrane component of the conventional exocytotic pathway which serves as a docking mechanism for classical exocytotic secretory vesicles. These observations have generated a working hypothesis which assumes that FGF-1 utilizes the cytosolic face of the conventional exocytotic pathway to gain access to a novel secretion pathway which employees a protease to cleave stg-1 free of its transmembrane support. Thus, the goal of this application is to confirm and expand their preliminary data that FGF-1 is secreted as a FGF-1 homodimer:stg-1 complex and to determine the structural basis for the interaction between FGF-1 and stg-1 including the identification of the stg-1-cleaving enzyme. They anticipate that this effort will not only elucidate a novel secretion pathway, but will also yield new insight into the mechanism by which FGF-1 modifies human physiologic and pathophysiologic states.

描述（改编自研究者摘要）：肝素结合 成纤维细胞生长因子（FGF）-1是一种有效的血管生成调节剂， 体内的神经营养现象，其通过包括 高亲和力FGF受体（R）家族的酪氨酸磷酸化 靶细胞的表面。 因为FGF-1不包含经典的 信号肽序列引导其通过内质网分泌 虽然它是网状高尔基体的一部分，但功能需要FGFR信号传导， 了解FGF-1的分泌机制很重要。 他们有 先前表明FGF-1是作为生物学上无活性和不良的生物学活性物质释放的， 肝素结合蛋白对体外温度应激的反应。 该实验室的其他研究表明，FGF-1（i）可以 被还原型谷胱甘肽激活，（ii）作为cys 30-cys 30 FGF-1释放 同二聚体，（iii）能够与磷脂酰丝氨酸结合， 亲和力，和（iv）与p65的胞质p45片段复合 突触结合蛋白（stg）-1在温度条件培养基和绵羊中 个脑袋 有趣的是，p65 stg-1是一种跨膜成分， 传统的胞吐途径，其作为对接机制， 典型的胞吐分泌囊泡。 这些观察产生了 一个工作假设，假设FGF-1利用的胞质面的 传统的胞吐途径获得一种新的分泌物 利用蛋白酶切割STG-1使其跨膜 支持. 因此，本申请的目标是确认和扩展其 FGF-1作为FGF-1同源二聚体：stg-1复合物分泌的初步数据 并确定FGF-1和TGF-β 1之间相互作用的结构基础。 STG-1，包括STG-1裂解酶的鉴定。 他们 预计这项工作不仅将阐明一种新的分泌物， 途径，但也将产生新的见解的机制，FGF-1 改变人类生理和病理生理状态。