Dongwan Cheng, Li Zheng, Junjie Hou, Jifeng Wang, Peng Xue, Fuquan Yang, Tao Xu. A new dimethyl labeling-based SID-MRM-MS method and its application to three proteases involved in insulin maturation. Biophysics Reports, 2015, 1(2): 71-80. doi: 10.1007/s41048-015-0012-1
Citation:
Dongwan Cheng, Li Zheng, Junjie Hou, Jifeng Wang, Peng Xue, Fuquan Yang, Tao Xu. A new dimethyl labeling-based SID-MRM-MS method and its application to three proteases involved in insulin maturation. Biophysics Reports, 2015, 1(2): 71-80. doi: 10.1007/s41048-015-0012-1
Dongwan Cheng, Li Zheng, Junjie Hou, Jifeng Wang, Peng Xue, Fuquan Yang, Tao Xu. A new dimethyl labeling-based SID-MRM-MS method and its application to three proteases involved in insulin maturation. Biophysics Reports, 2015, 1(2): 71-80. doi: 10.1007/s41048-015-0012-1
Citation:
Dongwan Cheng, Li Zheng, Junjie Hou, Jifeng Wang, Peng Xue, Fuquan Yang, Tao Xu. A new dimethyl labeling-based SID-MRM-MS method and its application to three proteases involved in insulin maturation. Biophysics Reports, 2015, 1(2): 71-80. doi: 10.1007/s41048-015-0012-1
The absolute quantification of target proteins in proteomics involves stable isotope dilution coupled with multiple reactions monitoring mass spectrometry (SID-MRM-MS). The successful preparation of stable isotope-labeled internal standard peptides is an important prerequisite for the SID-MRM absolute quantification methods. Dimethyl labeling has been widely used in relative quantitative proteomics and it is fast, simple, reliable, cost-effective, and applicable to any protein sample, making it an ideal candidate method for the preparation of stable isotope-labeled internal standards. MRM mass spectrometry is of high sensitivity, specificity, and throughput characteristics and can quantify multiple proteins simultaneously, including low-abundance proteins in precious samples such as pancreatic islets. In this study, a new method for the absolute quantification of three proteases involved in insulin maturation, namely PC1/3, PC2 and CPE, was developed by coupling a stable isotope dimethyl labeling strategy for internal standard peptide preparation with SID-MRM-MS quantitative technology. This method offers a new and effective approach for deep understanding of the functional status of pancreatic β cells and pathogenesis in diabetes.
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