DYNAMIC SILAC FOR THE STUDY OF PRION PROPAGATION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 Prion是一种感染性蛋白质聚集体，会导致哺乳动物的神经退化和死亡。培养的神经细胞已被用于研究普恩病毒。在这些细胞系中，由于不能区分提供给启动感染的新的细胞内的普恩，所以对普恩的传播研究变得复杂起来。将一种新的表位引入到细胞内的Pron蛋白中已经被用来确定Pron的来源。这项技术是有问题的，因为蛋白质结构的扰动可能会影响Pron的传递。培养基中标记的氨基酸将被加入到新合成的普鲁恩中。同位素标记与质谱学相结合，将使我们能够识别细胞培养中产生的普恩。 与加州大学旧金山分校质谱学设施的合作将使我们能够获得必要的仪器，以测量我们产生的普里子中的同位素掺入程度。我们已经在类似的项目上进行了成功的合作。我们设想这个项目将使用我们之前建立的协议。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Prions are infectious protein aggregates which cause neurodegeneration and death in mammals. Cultured neuronal cells have been used in the study of prions. The study of prion propagation in these cell lines is complicated by an inability to differentiate prions supplied to initiate the infection from new cellular prions. Introducing an novel epitope into cellular prion protein has been used previously to identify prion origin. This technique is problematic because pertubations to protein structure may influence prion transmission. Labeled amino acids in the culture medium would be incorporated into newly synthesized prions. The isotopic label in combination with mass spectroscopy would allow us to identify prions produced in the cell culture. Collaboration with the UCSF Mass Spectrometry Facility would give us access to the instrumentation necessary to measure the extent of isotopic incorporation into the prions we have generated. We have had successful collaborations on similar projects. We envision that this project would use the protocols we have established previously.