Sköld Karl, Svensson Marcus, Nilsson Anna, Zhang Xiaoqun, Nydahl Katarina, Caprioli Richard M, Svenningsson Per, Andrén Per E
Laboratory for Biological and Medical Mass Spectrometry, Uppsala University, Box 583 Biomedical Centre, SE-75123 Uppsala, Sweden.
J Proteome Res. 2006 Feb;5(2):262-9. doi: 10.1021/pr050281f.
PEP-19 is a neuronal calmodulin-binding protein, and as such, a putative modulator of calcium regulated processes. In the present study, we used proteomics technology approaches such as peptidomics and imaging MALDI mass spectrometry, as well as traditional techniques (immunoblotting and in situ hybridization) to identify PEP-19 and, specifically, to measure PEP-19 mRNA and protein levels in an animal model of Parkinson's disease. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration in mice resulted in a significant decrease in striatal PEP-19 mRNA. Capillary nano-flow liquid chromatography electrospray mass spectrometry analysis of striatal tissue revealed a significant decrease of the PEP-19 protein level. Moreover, imaging MALDI mass spectrometry also showed that PEP-19 protein was predominantly localized to the striatum of the brain tissue cross sections. After MPTP administration, PEP-19 levels were significantly reduced by 30%. We conclude that PEP-19 mRNA and protein expression are decreased in the striatum of a common animal model of Parkinson's disease. Further studies are needed to show the specific involvement of PEP-19 in the neurodegeneration seen in MPTP lesioned animals. Finally, this study has shown that the combination of traditional molecular biology techniques with novel, highly specific and sensitive mass spectrometry methods is advantageous in characterizing molecular events of many diseases, including Parkinson's disease.
PEP - 19是一种神经元钙调蛋白结合蛋白，因此是钙调节过程的一种假定调节剂。在本研究中，我们使用了蛋白质组学技术方法，如肽组学和成像基质辅助激光解吸电离质谱，以及传统技术（免疫印迹和原位杂交）来鉴定PEP - 19，特别是测量帕金森病动物模型中PEP - 19的mRNA和蛋白质水平。给小鼠注射1 - 甲基 - 4 - 苯基 - 1,2,3,6 - 四氢吡啶（MPTP）导致纹状体中PEP - 19 mRNA显著降低。对纹状体组织进行毛细管纳流液相色谱电喷雾质谱分析显示PEP - 19蛋白水平显著降低。此外，成像基质辅助激光解吸电离质谱还表明PEP - 19蛋白主要定位于脑组织横截面的纹状体。注射MPTP后，PEP - 19水平显著降低了30%。我们得出结论，在常见的帕金森病动物模型的纹状体中，PEP - 19的mRNA和蛋白表达降低。需要进一步研究以表明PEP - 19在MPTP损伤动物中所见神经退行性变中的具体作用。最后，本研究表明，传统分子生物学技术与新颖、高度特异性和灵敏的质谱方法相结合，在表征包括帕金森病在内的许多疾病的分子事件方面具有优势。
