IDENTIFICATION AND CHARACTERIZATION OF BIOMARKERS OF ORGANOPHOSPHORUS EXPOSURES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 全球每年生产超过10亿磅的有机磷(OP)化学品，其中包括在美国喷洒的7000万磅杀虫剂。目前监测毒死蜱等有机磷农药环境和职业暴露的方法具有局限性，包括特异性和敏感度低，检测时间窗口短。OP三甲基磷酸盐(TCP)可污染喷气发动机排出的空气，并导致商业航空公司飞行员、机组人员和乘客的职业暴露，其生物标志物尚未确定。我们工作的目的是识别、提纯和表征新的OP暴露生物标志物。丁酰胆碱酯酶(BChE)抑制已成为监测OP暴露的标准。通过识别和表征半衰期更长的分子生物标志物，我们应该能够在临床上检测到比目前可能的更长时间的TCP和OP杀虫剂的暴露。酰基肽水解酶(APH)是一种红细胞(RBC)胞浆丝氨酸蛋白酶，它能从多肽中去除N-乙酰化氨基酸，并裂解氧化蛋白。由于其特性，它是一个极好的暴露生物标记物。我们已经能够提纯APH，并检测到CPS和TCP的代谢产物对APH的抑制作用。RBC的120天寿命为检测暴露提供了一个更长的窗口。本研究利用功能蛋白质组学和酶活性来鉴定和表征神经毒性、环境和职业性OP暴露的有用生物标志物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Over 1 billion pounds of organophosphorus (OP) chemicals are manufactured worldwide each year, including 70 million pounds of pesticides sprayed in the US. Current methods to monitor environmental and occupational exposures to OPs such as chlorpyrifos (CPS) have limitations, including low specificity and sensitivity, and short time windows for detection. Biomarkers for the OP tricresyl phosphate (TCP), which can contaminate bleed air from jet engines and cause an occupational exposure of commercial airline pilots, crewmembers and passengers, have not been identified.The aim of our work has been to identify, purify, and characterize new biomarkers of OP exposure. Butyrylcholinesterase (BChE) inhibition has been a standard for monitoring OP exposure. By identifying and characterizing molecular biomarkers with longer half-lives, we should be able to clinically detect TCP and OP insecticide exposure after longer durations of time than are currently possible.Acylpeptide hydrolase (APH) is a red blood cell (RBC) cytosolic serine proteinase that removes N-acetylated amino acids from peptides and cleaves oxidized proteins. Due to its properties, it is an excellent candidate for a biomarker of exposure. We have been able to purify APH and detect inhibition by both CPS and metabolites of TCP. The 120-day lifetime of the RBC offers a much longer window for detecting exposure. The OP-modified serine conjugate in the active site tryptic peptide has been characterized by mass spectrometry.This research uses functional proteomics and enzyme activities to identify and characterize useful biomarkers of neurotoxic environmental and occupational OP exposures.