REGULATION OF NEUROPEPTIDE BIOSYNTHESIS

神经肽生物合成的调控

• 批准号: 3226164
• 负责人: JACK E DIXON
• 金额: $ 25.48万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-10-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3226164
• 关键词: carboxypeptidase; gene expression; genetic manipulation; genetic regulation; genetic translation; laboratory rabbit; laboratory rat; neuropeptide Y; neuropeptides; neurotransmitter biosynthesis; neurotrophic factors; nucleic acid sequence; posttranslational modifications; protein sequence; transcription factor

The long range objective of this project is to apply the techniques of biochemistry to investigate how genes of the nervous system are expressed and regulated. The nervous system is composed of many neurons which interconnect with one another in a precis manner. The diversity of neurons and the nature of their interactions suggest that specific genes or sets of genes are activated in some neurons and not in others. One of the specific aims of this proposal is to develop a better understanding of the molecular events associated with the expression and regulation of the most abundant and widely distributed neuropeptide the mammalian nervous system, neuropeptide Y. The specific regions of the NPY gene responsible for expression, tissue-specific expression, and response to nerve growth factor will be dissected using molecular and biochemical approaches. Most neuropeptides and numerous proteins undergo post-translational processing which requires limited proteolysis. At least two (and often more) enzymes appear to function in their processing: 1) a trypsin-like protease, which recognizes precursors to both proteins and neuropeptides where two adjacent basic residues are present; and 2) a carboxypeptidase B-like enzyme, which removes the C-terminal basic residues form the C- terminus of intermediates which are formed by limited proteolysis. These second aim of this proposal is to better define the molecular events associated with the limited proteolysis of precursors to proteins and neuropeptides. Our laboratory has identified and sequenced a full-length clone of the carboxypeptidase B-like processing enzyme (referred to as carboxypeptidase H [CPH]). We have shown that carboxypeptidase H is synthesized as a precursor, which is the first reported example of a proteolytic-processing enzyme being synthesized as a precursor. The identification and characterization of the intermediates produced during the processing of this precursor is also a goal of this work. In addition, the gene encoding carboxypeptidase H should contain sequences which result in its selective expression in hormone producing cells. Therefore, we would like to identify and characterize these regions of the gene.

该项目的长期目标是应用以下技术： 生物化学研究神经系统的基因如何表达 和监管。 神经系统由许多神经元组成， 以精确方式彼此互连。 之多样 神经元及其相互作用的性质表明， 或者基因组在某些神经元中被激活，而在另一些神经元中则没有。 之一 这项建议的具体目的是为了更好地了解 的表达和调节相关的分子事件， 哺乳动物中最丰富和分布最广的神经肽 神经系统，神经肽Y。 NPY基因的特定区域 负责表达、组织特异性表达和对 神经生长因子将使用分子和生物化学方法进行解剖， 接近。 大多数神经肽和许多蛋白质都经历翻译后 这需要有限的蛋白水解。至少两个（而且经常 更多）酶似乎在其加工过程中起作用：1）胰蛋白酶样 蛋白酶，识别蛋白质和神经肽的前体 其中存在两个相邻的碱性残基;和2）羧肽酶 类B酶，其从C-末端除去C-末端碱性残基， 通过有限的蛋白水解形成的中间体的末端。这些 该建议的第二个目的是更好地定义分子事件 与蛋白质前体的有限蛋白水解有关， 神经肽 我们的实验室已经鉴定并测序了一个全长的 羧肽酶B样加工酶（称为羧肽酶B样加工酶）的克隆 羧肽酶H [CPH]）。 我们已经证明羧肽酶H是 作为前体合成，这是第一个报道的 蛋白水解加工酶作为前体被合成。 的 中间体的鉴别和表征 这种前体的加工也是这项工作的目标。 在 此外，编码羧肽酶H的基因应含有序列 这导致其在激素产生细胞中的选择性表达。 因此，我们想确定和描述这些区域的特征， 基因