PUROMYCIN SENSITIVE (ENKEPHALIN) AMINOPEPTIDASE ISOFORMS

这项研究的目的是研究表达和 调节嘌呤霉素敏感的氨基肽酶（脑啡肽- 降解氨肽酶），其已显示降解神经肽 包括脑啡肽和强啡肽。最近，三种异构体 该酶是从cDNA文库中分离的。我们将利用这些 cDNA以分离酶的基因组克隆，从而确定酶的来源 这些异构体。预期这三种同种型来自于 单个基因，因为它们的大多数编码几乎相同 序列的将对基因组克隆的外显子进行测序，以确定 它们的拼接模式我们将确定酶是如何调节的， 细胞，包括每个同种型在细胞中的位置，无论 并非每种异构体都具有催化活性，以及这些异构体是如何 在空间和时间上表达。最后，转基因小鼠将 用于研究酶同工型对发育的假定作用 以及对细胞周期的调节。

The objective of this research proposal is to study the expression and regulation of the puromycin-sensitive aminopeptidase (enkephalin- degrading aminopeptidase), which has been shown to degrade neuropeptides including the enkephalins and dynorphins. Recently, three isoforms of this enzyme were isolated from a cDNA library. We will utilize these cDNAs to isolate a genomic clone(s) of the enzyme to determine the origin of these isoforms. It is expected that the three isoforms arise from a single gene due to the near identity of the majority of their coding sequences. Exons of the genomic clone will be sequenced to determine their splicing pattern. We will determine how the enzyme is regulated in cells, including where each isoform is located in the cell, whether or not each isoform is catalytically active, and how these isoforms are expressed spatially and temporally. Finally, transgenic mice will be employed to study the putative role of the enzyme isoforms on development and on regulation of the cell cycle.