PUROMYCIN SENSITIVE (ENKEPHALIN) AMINOPEPTIDASE ISOFORMS

这项研究计划的目的是研究表达和 对嘌呤霉素敏感的氨基肽酶(脑啡肽)的调节 降解氨基肽酶)，它已被证明能降解神经肽 包括脑啡肽和强啡肽。最近，三种亚型的亚型 该酶是从一个cDNA库中分离得到的。我们将利用这些 从DNA中分离出基因组克隆的酶(S)，确定其来源 这些同工异构体。据推测，这三种异构体来自一种 单个基因由于其大部分编码的近乎相同 序列。基因组克隆的外显子将被测序以确定 它们的拼接模式。我们将确定这种酶是如何在 细胞，包括每个异构体在细胞中的位置，无论是或 不是每种异构体都具有催化活性，以及这些异构体是如何 在空间和时间上表现出来的。最后，转基因小鼠将被 用来研究酶异构体在发育中的假定作用 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.