BIOSYNTHESIS AND PROCESSING OF PEPTIDE HORMONE

描述：（改编自申请人的摘要）激素生物合成 需要选择性地将激素包装到分泌囊泡中。授出 着重于分泌囊泡从高尔基体网络的萌芽 (TGN)磷脂代谢和蛋白质因子在其中的作用 发芽最终目标包括合理的药物设计，以治疗病理性 激素分泌的条件。正在推行的基本计划包括： ADP-核糖基化因子-1（ARF-1）激活TGN中的磷脂酶D。 然后磷脂酶D（PLD）又产生磷脂酸（PA）。目标1： PA积累在调节磷脂酰肌醇生物合成中的作用 将检查4，5二磷酸盐（PIP 2）。假设PA调节 PIP 2，控制TGN的出芽和高尔基体的维持 架构目标2：目标1将扩展到其他细胞，看看有哪些方面 激素分泌、高尔基体形态、组成性分泌和溶酶体 囊泡的释放取决于PA的产生。目的3：蛋白质的纯化 促进囊泡出芽的因子，将使用标准蛋白质进行研究。 纯化方法。假设PLD被招募到TGN， 通过由ARF-1和其他蛋白质组成的蛋白质复合物激活。原生 ARF-1的表位标记形式将与 交联剂以鉴定所提出的蛋白质复合物。

DESCRIPTION: (Adapted from the applicant's abstract) Hormone biosynthesis requires the selective packaging of hormones into secretory vesicles. The grant focuses on the budding of secretory vesicles from the trans Golgi network (TGN), and the roles of phospholipid metabolism and protein factor in that budding. Eventual goals include rational drug design to treat pathological conditions of hormone secretion. The basic scheme being pursued involves activation of phospholipase D in the TGN by ADP-ribosylation factor-1 (ARF-1). Then phospholipase D (PLD) in turn generates phosphatidic acid (PA). Aim 1: The role of PA accumulation in regulating the biosynthesis of phosphatidylinositol 4,5 bisphosphate (PIP2) will be examined. The hypothesis is that PA regulates PIP2, which controls budding from the TGN and also maintenance of Golgi architecture. Aim 2: Aim 1 will be extended to other cells to see what aspects of hormone secretion, Golgi morphology, constitutive secretion, and lysosomal vesicle release depends on PA production. Aim 3: Purification of protein factors, which promote vesicle budding, will be pursued using standard protein purification approaches. The hypothesis is that PLD is recruited to the TGN and activated via a protein complex consisting of ARF-1 and other proteins. Native and epitope tagged versions of ARF-1 will be used in conjunction with cross-linking agents to identify the proposed protein complex.