OPIOID PEPTIDE PROCESSING ENZYMES

描述(申请人摘要)： 请求续签K02奖励是为了使申请者免于 实质性教学和行政责任要增加 研究生产力。在过去的4年里，私家侦探能够 将80%以上的时间投入到研究活动中，这是一种努力 这导致了显著增强的执行能力 实验，为博士后研究员和研究生提供建议，并获得 通过协作和亲身实践实现的新技术 实验。这一研究工作的加强预计将 在应用程序续订期间继续，并受保护 由K02奖提供；如果没有它，教学责任很可能 急剧增加，当然最显著的是通过假设 在一个主要的服务课程中担任董事。 该提案基于目前正在进行的两个项目，其中一个是 阿片肽前体的蛋白水解性加工，以及一种 PC2结合蛋白7B2的作用机制。四个具体目标 1)确定重组PC1和PC2的特异性 重组正常和突变的前脑啡肽原及其相关的荧光 底物；这些实验将描绘出底物偏好 两台电脑。2)寻找新的PC1抑制剂，研究其生物化学和生物活性。 PC1和PC2抑制剂的细胞生物学，并评估其潜在作用 体内脑啡肽原加工的靶向抑制剂。这些实验 将补充关于酶的第一个具体目标中描述的那些 关于合成酶设计的专一性和应产生的信息 抑制剂。3)确定7B2在ProPC2调控中的作用 神经内分泌细胞系的转化，重点是细胞的命运 前肽及其与7B2裂解/解离的关系 表格。4)探讨7B2和PC2的分子相互作用。 体外实验。综上所述，这些实验应该有助于我们 了解阿片类药物前激素转换酶的基本生物化学 多肽前体以及转换酶活性的调节。

DESCRIPTION (Applicant's Abstract): This renewal of a K02 award is requested in order to free the applicant from substantive teaching and administrative responsibilities to increase research productivity. During the past 4 years the PI has been able to devote more than 80% of her time to research activities, a level of effort which has resulted in a significantly enhanced ability to perform experiments, advise postdoctoral fellows and graduate students, and acquire new technologies both by collaboration as well as by hands-on experimentation. This enhancement of research effort is expected to continue during the renewal period of the application with the protection afforded by the K02 award; without it, teaching responsibilities are likely to increase dramatically, most notably by the assumption of course directorship in a major service course. The proposal is based upon two currently ongoing projects, one on the proteolytic processing of opioid peptide precursors, and one on the mechanism of action of the PC2 binding protein 7B2. The four specific aims are 1) to define the specificity of recombinant PC1 and PC2 using recombinant normal and mutant proenkephalins and related fluorogenic substrates; these experiments will delineate substrate preferences of the two PCs. 2) To identify novel PC1 inhibitors, study the biochemistry and cell biology of PC1 and PC2 inhibitors, and assess potential effects of targeted inhibitors on proenkephalin processing in vivo. These experiments will complement those described in the first specific aim as to enzyme specificity and should yield information on the design of synthetic enzyme inhibitors. 3) To determine the role of 7B2 in the regulation of proPC2 conversion in neuroendocrine cel1 lines, with a focus on the fate of the propeptide and the relationship of its cleavage/dissociation to that of 7B2 forms. 4) To explore the molecular interaction of 7B2 and PC2 through in vitro experiments. Taken together, these experiments should help us to understand the basic biochemistry of the prohormone convertases on opioid peptide precursors as well as the regulation of convertase activity.