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: 8713452
• 负责人: Thomas Silhavy
• 金额: $ 19.08万
• 依托单位: Princeton University
• 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 1991-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8713452&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 energy 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.

这是细菌遗传学家(西尔哈维博士)和物理化学家(吉拉施博士)合作项目的一部分，目的是研究信号序列在蛋白质输出中的分子功能。大多数出口蛋白质最初是以带有氨基末端信号序列的前体形式合成的。已经提出了令人信服的证据，表明这一序列在出口过程中的重要作用。然而，其功能的细节(S)并不是很清楚。由于其体积小(15-30个氨基酸)，这种输出信号特别适合使用现代遗传和物理技术进行分析，例如指定杂交蛋白质的基因融合、使用合成寡核苷酸的定点突变、固态肽合成以及模型膜系统中的荧光能量转移和猝灭。这些研究人员的实验室以前的结果表明，信号序列的内在物理化学性质与其在体内的功能相关。该项目将扩展和放大正在进行的跨学科方法，以剖析出口蛋白质的信号序列与生物膜相互作用的分子机制。这一目标将通过使用出口缺陷的大肠杆菌突变株来促进，在该突变株中，外膜蛋白Lamb的信号序列被改变，并通过相应的合成信号肽的物理化学研究。