Proteomics in neurotoxicology

我们正在应用蛋白质组学方法解决神经病理疾病中尚未解决的问题。识别神经精神疾病的蛋白质生物标志物的方法，如小儿患者链球菌感染后的强迫症（PANDAS），正在设计和测试中。免疫亲和策略的组合用于从患者血清中分离蛋白质。正在比较蛋白质的质谱模式，以确定是否存在患者状态和疾病特征的统计显著特征。我们制备了几个亚蛋白质组，并应用化学计量学分析来测试完整蛋白质的MALDI/TOF测量是否产生疾病特征图谱。第二种策略是从基因组研究中分离出与精神分裂症相关的蛋白质。Dysbindin抗体的免疫亲和力正在用现有的基因失调基因修饰的小鼠品系进行测试。

We are applying proteomic methodology to unresolved problems in neuropathologic diseases. Methods to identify protein biomarkers of neuropsychiatric disorders, such as the obsessive compulsive syndrome that follows streptococcal infections (PANDAS) in pediatric patients, are being devised and tested. A combination of immunoaffinity strategies is used to separate proteins from patient sera. Mass spectral patterns of proteins are being compared to determine whether there are statistically significant characteristics of patient state and the disease trait. We prepared several sub-proteome fractions and have applied chemometric analyses to test whether MALDI/TOF measurements of the intact proteins produce disease characteristic profiles. A second strategy is to isolate proteins implicated from genomic studies as being associated with schizophrenia. Dysbindin antibody immunoaffinity is being tested with available genetically dysbindin-gene modified mouse strains. In collaborative studies with NHGRI, the mitochondrial electron transport complex-I proteomics are being characterized. The hypothesis that deficiency in this complex results in Parkinson?s Disease is being tested. Elucidation of the complexity of a protein?s function and its role in biological pathways involves first identifying potential protein-protein interactions. Comparison of these interactions for wild-type vs. modified protein forms with a biologically relevant complex protein mixture can yield quantitative differences in the extent of these interactions relative to its stage in the cellular pathway. The standard methods of pull down assays combined with solution digestion, LC-MS/MS and data base searching provide for a high-throughput strategy for the identification differences in protein-protein interactions. Equally useful are the quantitative differences/similarities imbedded in the measured ion currents for specific peptides. Much attention has been given to the use of stable isotope labeling for relative quantization of protein interactions. The stability of high-throughput LC-MS/MS ion currents are such that integration of ion current alone can be used for rapid screening of up/down regulated proteins occurring between multiple samples. In collaborative studies with NYU, we are performing proteome analyses of several affinity purified protein extracts from HeLa cells expressing the amino terminus of the Huntington disease protein with different numbers of glutamines (the wild type and disease states). The objective of this project is to test the hypothesis that there is transcriptional dysregulation associated with Huntington's Disease.