Sprenger Adrian, Küttner Victoria, Bruckner-Tuderman Leena, Dengjel Jörn
Freiburg Institute for Advanced Studies, School of Life Sciences, Center for Biological Systems Analysis, University of Freiburg, Freiburg, Germany.
Methods Mol Biol. 2013;961:179-91. doi: 10.1007/978-1-62703-227-8_10.
Physiological functions of skin cells are often altered in diseases. Since the underlying molecular mechanisms are generally executed by proteins, it is of interest to assess protein dynamics in normal and pathologically altered cells. These can be readily analyzed in relevant cell culture models by quantitative mass spectrometry (MS)-based proteomics, which is the method of choice to track the concerted action and spatial relocation of unknown involved factors in an unbiased way. Different quantitative MS strategies have been used to characterize protein dynamics. In this chapter we describe in detail the use of stable isotope labeling by amino acids in cell culture for an unbiased quantitative analysis of protein dynamics in the two major cell types of the skin, keratinocytes and fibroblasts.
皮肤细胞的生理功能在疾病中常常发生改变。由于潜在的分子机制通常由蛋白质执行，因此评估正常细胞和病理改变细胞中的蛋白质动态变化具有重要意义。通过基于定量质谱（MS）的蛋白质组学，可以在相关细胞培养模型中轻松分析这些变化，这是一种以无偏倚方式追踪未知相关因子协同作用和空间重定位的首选方法。不同的定量质谱策略已被用于表征蛋白质动态变化。在本章中，我们将详细描述细胞培养中氨基酸稳定同位素标记的应用，用于对皮肤的两种主要细胞类型——角质形成细胞和成纤维细胞中的蛋白质动态变化进行无偏倚定量分析。
