Wieseler Stacy, Schopfer Lawrence, Lockridge Oksana
University of Nebraska Medical Center, Omaha, NE 68198-6805, USA.
J Mol Neurosci. 2006;30(1-2):93-4. doi: 10.1385/JMN:30:1:93.
The goal of this work is to identify novel serum proteins that are labeled with organophosphates (OPs) and to create a protocol for identification using mass spectroscopy. The use of OP-labeled proteins for identification of exposure is useful because such proteins will remain in circulation for weeks (Van Der Schans et al., 2004). Currently, both butyrylcholinesterase (BChE) and albumin have been shown to bind OPs in blood. Peeples et al. (2005) showed that albumin is labeled by OPs, specifically 6-Nbiotinylaminohexyl isopropyl phosphorofluoridate hemihydrate, in living mice. Albumin is the major protein in human serum, and its reaction with OPs tends to overwhelm the identification of other proteins. In vitro studies of human serum require removal of the serum albumin without depleting the less abundant proteins. Following this step, identifying the remaining proteins is simply a matter of labeling the proteins with an OP, separating the labeled from nonlabeled proteins, and using Q-trap mass spectrometry for identification.
这项工作的目标是鉴定用有机磷酸酯(OPs)标记的新型血清蛋白,并创建一种使用质谱进行鉴定的方案。使用OP标记的蛋白来鉴定暴露情况是有用的,因为这类蛋白会在血液循环中留存数周(范德尚斯等人,2004年)。目前,已证实血液中的丁酰胆碱酯酶(BChE)和白蛋白都会与OPs结合。皮普尔斯等人(2005年)表明,在活体小鼠中,白蛋白会被OPs标记,特别是6-生物素氨基己基异丙基磷酰氟半水合物。白蛋白是人类血清中的主要蛋白质,它与OPs的反应往往会掩盖其他蛋白质的鉴定。对人类血清进行的体外研究需要去除血清白蛋白,同时又不耗尽含量较少的蛋白质。在这一步之后,鉴定剩余的蛋白质只需用一种OP标记这些蛋白质,将标记的与未标记的蛋白质分离,然后使用Q-阱质谱进行鉴定。