BIOSYNTHESIS AND PROCESSING OF PEPTIDE HORMONE

描述：(改编自申请者摘要)荷尔蒙生物合成 需要选择性地将荷尔蒙包装成分泌囊。助学金 重点研究了来自跨高尔基体网络的分泌囊泡的萌发 (TGN)，以及磷脂代谢和蛋白质因子在其中的作用 萌芽中。最终目标包括合理设计治疗病理性疾病的药物 激素分泌的条件。正在推行的基本计划包括 ADP-核糖化因子-1(ARF-1)激活TGN中的磷脂酶D。 然后，磷脂酶D(PLD)反过来产生磷脂酸(PA)。目标1： PA蓄积在调节磷脂酰肌醇生物合成中的作用 4，5-二磷酸(PIP2)将被检测。假设是PA调节 PIP2，控制来自TGN的萌发以及高尔基体的维护 建筑。目标2：目标1将扩展到其他单元格，以了解哪些方面 激素分泌、高尔基体形态、结构性分泌和溶酶体 囊泡的释放取决于PA的产生。目标3：蛋白质的纯化 促进囊泡萌发的因素将使用标准蛋白质进行研究。 提纯方法接近了。假设PLD被招募到TGN并 通过由ARF-1和其他蛋白质组成的蛋白质复合体激活。原生的 ARF-1的表位标记版本将与 用交联剂来鉴定提议的蛋白质复合体。

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.