Enzymic Detoxification for Organophosphate Nerve Agents

有机磷神经毒剂的酶解毒

基本信息

批准号：
7659598
负责人：
Frank M. Raushel
金额：
$ 30.18万
依托单位：
TEXAS A&M UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659598
关键词：
Active Sites Agriculture Amino Acid Sequence Chemical Weapons Cholinesterases Collaborations Decontamination Detection Diffusion Drug Metabolic Detoxication Engineering Enzymes Escherichia coli Esters Health Human Hydrolysis Insecticides Left Libraries Measurement Methods Modification Mutagenesis Organophosphates Paraoxon Phosphoric Triester Hydrolases Poison Procedures Property Proteins Protocols documentation Reporting Research Resolution Sarin Screening procedure Site Societies Soman Spectrum Analysis Stereoisomer Structure System Techniques Variant X ray diffraction analysis X-Ray Crystallography X-Ray Diffraction analog base catalyst combinatorial cyclosarin design design and construction directed evolution enzyme activity enzyme structure high throughput screening inorganic phosphate molecular recognition mutant nerve agent neurotransmission novel public health relevance tabun three dimensional structure

项目摘要

DESCRIPTION (provided by applicant): Organophosphate and organophosphonate esters are among the most toxic compounds that have ever been synthesized. These compounds are toxic because of their inherent ability to interfere with nerve transmission through the rapid and irreversible inactivation of the enzyme acetyl cholinesterase. The toxic properties of the organophosphates have been exploited as agricultural insecticides and as chemical weapons. A bacterial phosphotriesterase enzyme has been identified that can function as a catalyst for the recognition, hydrolysis, and detoxification of a broad spectrum of organophosphate nerve agents. The primary objective of this application is to design and characterize modified forms of the bacterial phosphotriesterase that are optimized for the selective recognition and destruction of those organophosphates that pose the most serious threats to human health. Rational and combinatorial libraries of mutant enzymes will be constructed and those variants with enhanced catalytic proficiency will be identified through novel screening and selection procedures. The progressive changes in the amino acid sequence of the bacterial phosphotriesterase will be directly correlated with the enhancements in the catalytic constants and the modifications within the active site as determined by X-ray diffraction methods. PUBLIC HEALTH RELEVANCE: The toxic properties of the organophosphate nerve agents represent a serious threat to the health and well being of civilized societies. The primary objective of this project is to create novel catalytic enzymes that are optimized in their ability to detect, destroy and detoxify a broad spectrum of organophosphate nerve agents.

描述（由申请人提供）：有机磷酸酯和有机膦酸酯是迄今为止合成的毒性最强的化合物之一。这些化合物具有毒性，因为它们具有通过快速且不可逆地灭活乙酰胆碱酯酶来干扰神经传递的固有能力。有机磷酸酯的毒性已被用作农业杀虫剂和化学武器。已鉴定出一种细菌磷酸三酯酶，可以作为广谱有机磷酸酯神经毒剂的识别、水解和解毒的催化剂。该应用的主要目标是设计和表征细菌磷酸三酯酶的修饰形式，该酶经过优化，可选择性识别和破坏对人类健康构成最严重威胁的有机磷酸酯。将构建突变酶的合理组合文库，并通过新的筛选和选择程序鉴定那些具有增强催化能力的变体。细菌磷酸三酯酶氨基酸序列的渐进变化将与通过X射线衍射方法测定的催化常数的增强和活性位点内的修饰直接相关。公众健康相关性：有机磷神经毒剂的毒性对文明社会的健康和福祉构成严重威胁。该项目的主要目标是创造新型催化酶，优化其检测、破坏和解毒广谱有机磷神经毒剂的能力。