The Application of SILAC Mouse in Human Body Fluid Proteomics Analysis Reveals Protein Patterns Associated with IgA NephropathyReportar como inadecuado




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Evidence-Based Complementary and Alternative MedicineVolume 2013 2013, Article ID 275390, 10 pages

Research Article

Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Science, Chinese Academy of Sciences, Shanghai 200031, China

Department of Nephrology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 725 Wanping Road, Shanghai 200032, China

Cardiovascular Centre, Department of Internal Medicine, University of Michigan Medical Centre, Ann Arbor, MI 48109, USA

Received 31 December 2012; Accepted 18 April 2013

Academic Editor: Aiping Lu

Copyright © 2013 Shilin Zhao et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Body fluid proteome is the most informative proteome from a medical viewpoint. But the lack of accurate quantitation method for complicated body fluid limited its application in disease research and biomarker discovery. To address this problem, we introduced a novel strategy, in which SILAC-labeled mouse serum was used as internal standard for human serum and urine proteome analysis. The SILAC-labeled mouse serum was mixed with human serum and urine, and multidimensional separation coupled with tandem mass spectrometry IEF-LC-MS-MS analysis was performed. The shared peptides between two species were quantified by their SILAC pairs, and the human-only peptides were quantified by mouse peptides with coelution. The comparison for the results from two replicate experiments indicated the high repeatability of our strategy. Then the urine from Immunoglobulin A nephropathy patients treated and untreated was compared by this quantitation strategy. Fifty-three peptides were found to be significantly changed between two groups, including both known diagnostic markers for IgAN and novel candidates, such as Complement C3, Albumin, VDBP, ApoA,1 and IGFBP7. In conclusion, we have developed a practical and accurate quantitation strategy for comparison of complicated human body fluid proteome. The results from such strategy could provide potential disease-related biomarkers for evaluation of treatment.





Autor: Shilin Zhao, Rongxia Li, Xiaofan Cai, Wanjia Chen, Qingrun Li, Tao Xing, Wenjie Zhu, Y. Eugene Chen, Rong Zeng, and Yueyi

Fuente: https://www.hindawi.com/



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