Collaborative Research on the Molecular Functions of the Signal Sequence in Protein Export

蛋白质输出中信号序列的分子功能合作研究

• 批准号: 8710437
• 负责人: Lila Gierasch
• 金额: $ 1.93万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-15 至 1988-01-01
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8710437&HistoricalAwards=false
• 关键词: Collaborative; Research; Molecular; Functions; Signal

This is part of a collaborative project between a bacterial geneticist (Dr. Silhavy) and a physical chemist (Dr. Gierasch) to study the molecular functions of the signal sequence in protein export. Most exported proteins are synthesized initially in precursor form with an amino-terminal signal sequence. Convincing evidence demonstrating the essential role of this sequence in the export process has been presented. However, details of its function(s) are not well understood. Because of its small size (15-30 amino acids), this export signal is particularly amenable to analysis using modern genetic and physical techniques, such as gene fusions that specify hybrid proteins, site-specific mutagenesis using synthetic oligonucleotides, solid-state peptide synthesis, and fluorescence enregy transfer and quenching in model membrane systems. Previous results from the laboratories of these researchers demonstrated that intrinsic physical-chemical properties of the signal sequence are correlated with its in vivo function. This project will expand and amplify the ongoing interdisciplinary approach to dissect the molecular mechanism by which the signal sequence of exported proteins interacts with a biological membrane. This goal will be facilitated through the use of export-defective mutant strains of E. coli in which the signal sequence of the outer membrane protein, LamB, is altered, and through the physical-chemical study of corresponding synthetic signal peptides.

这是一个合作项目的一部分， 遗传学家（Silhavy博士）和物理化学家（Gierasch博士）， 研究蛋白质中信号序列的分子功能 导出. 大多数输出的蛋白质最初是在 具有氨基末端信号序列的前体形式。 令人信服的证据表明，这一重要作用 在出口过程中的顺序已经提出。 然而，在这方面， 其功能的细节还不清楚。 因为 由于其较小的尺寸（15-30个氨基酸），这种输出信号 特别适合于使用现代遗传学和 物理技术，如基因融合，指定杂交 蛋白质，使用合成的定点诱变 寡核苷酸、固态肽合成和荧光 模型膜系统中的能量传递和猝灭。 这些研究人员的实验室先前的结果 表明，固有的物理化学性质的 信号序列与其体内功能相关。 这 该项目将扩大和放大正在进行的跨学科 一种分析信号的分子机制的方法， 输出的蛋白质序列与生物 膜的 这一目标将通过使用 E.在大肠杆菌中， 外膜蛋白LamB的序列发生改变， 通过对相应的合成物的物理化学研究， 信号肽