MASS SPECTROMETRY OF OPIOID NEUROPEPTIDERGIC SYSTEMS

大鼠δ阿片受体（OP 1）将采用基质辅助激光解吸飞行时间质谱（MALDI-TOF MS）进行表征。MALDI-TOF MS提供了分析完整受体所需的高质量能力、检测受体所需的高检测灵敏度以及分析胰蛋白酶消化产生的未分辨肽混合物的能力，该混合物将用于肽图谱和OP表征（将使用其他酶和溴化氰）。研究完整的、完全后修饰的受体是重要的。与用于研究受体的其他方法相比，这种基于MS的方法提供了高灵敏度，快速分析和最佳特异性。研究OP所需的MALDI-TOF MS的几个操作特性将被优化，包括：样品分析的速度（MALDI-TOF可以以大约100个样品的速度在一个板上分析100个样品。每个样本1分钟;该速率对于肽图谱和OP表征是重要的）;检测灵敏度将提高到fmol/阿莫尔水平;分子特异性将通过使用源后衰变（PSD）来优化以获得胰蛋白酶肽的氨基酸序列;并优化测量OP 1分子量（MW）的质量准确度（0.1%水平）通过使用延迟提取（DE）和反射器（R）。

The rat delta opioid receptor (OP1) will be characterized with matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS). MALDI-TOF MS offers the high mass capability that is needed to analyze an intact receptor, the high detection sensitivity that is needed to detect a receptor, and the ability to analyze the mixture of unresolved peptides that is produced by trypsin digestion and that will be used for peptide mapping and for characterization of the OP (Other enzymes and cyanogen bromide will be used). It is important to study the intact, fully post- translationally modified receptor. Compared to the other methods that are used to study receptors, this MS-based method offers high sensitivity, rapid analysis, and optimum specificity. Several operating features of MALDI-TOF MS that are needed to study the OP will be optimized, including: the speed of sample analysis (MALDI-TOF can analyze 100 samples on a plate at a rate of ca. one minute per sample; that rate is important for peptide mapping and OP characterization); detection sensitivity will be improved to the fmol/amol level; molecular specificity will be optimized by using post-source decay (PSD) to obtain the amino acid sequence of tryptic peptides; and the mass accuracy of measuring the molecular weight (MW) of the OP1 will be optimized (0.1% level) by using delayed extraction (DE) and a reflectron (R).