MASS SPECTROMETRY ANALYSIS OF THE HUMAN CSF PROTEOME

描述：(改编自申请人的摘要)我们假设 从人脑脊液中获得的蛋白质组在对照组之间存在差异 那些被诊断为特发性下腰痛的患者是因为我们的 初步数据清楚地表明，这两个方面存在显著差异 组中特定阿片类药物和特定速激肽的量 神经肽。我们将扩大我们的研究范围，包括蛋白质和酶。我们会 首先从定性和定量两个方面对我们的假设进行实验验证 脑脊液中阿片和速激肽神经肽能系统的分析 蛋白质组，包括每个神经肽前体和相关的酶。 内啡肽和其他相关的神经肽系统也将被研究。这个 蛋白质组将用电泳法和质谱法进行分析。 我们将分析代谢级联反应中含有神经肽的蛋白质 合成神经元中的每个神经肽：DNA-和GT；RNA-和GT； 中等大小的蛋白质-神经肽-代谢物。这种连锁反应涉及到 几种不同的酶(激素原转换酶、氨基肽酶、肽 甘氨酸酰胺化单加氧酶、脑啡肽酶等)。 利多卡因的脊柱鉴别诊断很容易在 三个不同的腰背痛患者群体-对照(疼痛和非疼痛 患者)、生理应答者(需要手术)和非生理性应答者 响应者。非生理性应答者包括两个亚组：伪装者 和特发性腰背病人。伪装者很容易被识别(约95% 准确性)通过心理测试(MMPI)，很容易被排除在外 学习。我们的假设只关注特发性下腰痛，这一组 对他们的痛苦没有已知的原因。

DESCRIPTION: (adapted from applicant's abstract) We hypothesize that the proteome obtained from human cerebrospinal fluid (CSF) differs between controls and those patients who are diagnosed with idiopathic low back pain because our preliminary data clearly demonstrate significant differences in those two groups in the amount of a specific opioid and a specific tachykinin neuropeptide. We will expand our study to include proteins and enzymes. We will experimentally test our hypothesis by first qualitatively and quantitatively analyzing the opioid and tachykinin neuropeptidergic systems in the CSF proteome, including each neuropeptide precursor and associated enzymes. Endomorphins and other pertinent neuropeptide systems will also be studied. The proteome will be analyzed with electrophoresis and mass spectrometry. We will analyze the neuropeptide-containing proteins in the metabolic cascade that synthesizes each neuropeptide in the neuron: DNA -> RNA -> intermediate-sized proteins-> neuropeptide ->metabolites. That cascade involves several different enzymes (prohormone convertases, aminopeptidases, peptidyl glycine-amidating monooxygenase, enkephalinase, and others). A differential spinal diagnosis with lidocaine readily differentiates among the three different low back pain patient populations-controls (pain; non-pain patients), physiologic responders (require surgery), and nonphysiologic responders. The non-physiologic responders contain two subgroups: malingerers and idiopathic low back patients. Malingerers are readily identified (> ca. 95% accuracy) by a psychological test (MMPI), and are readily excluded from this study. Our hypothesis focuses only on the idiopathic low back pain group, which has no known reason for their pain.