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    標題: Comparative proteomics, network analysis and post-translational modification identification reveal differential profiles of plasma Con A-bound glycoprotein biomarkers in gastric cancer
    作者: Uen, Yih-Huei
    Lin, Kai-Yuan
    Sun, Ding-Ping
    Liao, Chen-Chung
    Hsieh, Ming-Song
    Huang, Yung-Kai
    Chen, Yen-Wei
    Huang, Pei-Hsuan
    Chen, Wei-Jung
    Tai, Chih-Chun
    Lee, Kuan-Wei
    Chen, You-Chia
    Lin, Ching-Yu
    貢獻者: 生物科技系
    關鍵字: Gastric Cancer
    Concanavalin A
    Mass Spectrometry
    Post-Translational Modification
    Human Plasma
    日期: 2013-05
    上傳時間: 2014-05-26 10:44:21 (UTC+8)
    出版者: Elsevier Science Bv
    摘要: In the study, we used Con A affinity chromatography, 1-D gel electrophoresis, and nano-LC-MS/MS to screen biomarker candidates in plasma samples obtained from 30 patients with gastric cancer and 30 healthy volunteers. First, we pooled plasma samples matched by age and sex. We identified 17 differentially expressed Con A-bound glycoproteins, including 10 upregulated proteins and 7 downregulated proteins; these differences were significant (Student's t-test, p-value < 0.05). Furthermore, 2 of the upregulated proteins displayed expression levels that were increased by 2-fold or more in gastric cancer samples when compared with normal control samples. These proteins included leucine-rich alpha-2-glycoprotein (LRG1) and inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3), and the expression levels were validated by Western blot analysis. Pathway and network analysis of the differentially expressed proteins by Ingenuity Pathway Analysis revealed vital canonical pathways involving acute phase response signaling, the complement system, LXR/RXR activation, hematopoiesis from pluripotent stem cells, and primary immunodeficiency signaling. Our results suggest that Con A-bound LRG1 and ITIH3 may not be practically applicable as a robust biomarker for the early detection of gastric cancer. Additionally, three novel PTMs in ITIH3 were identified and include hexose-N-acetyl-hexosamine at asparagine-(41), trimethylation at aspartic acid-(290), and flavin adenine dinucleotide at histidine-(335).Biological significanceOur study was to describe a combinatorial approach of Con A affinity chromatography, 1-D SDS-PAGE, and nano-LC/MS/MS that provides a label-free, comparative glycoproteomic quantification strategy for the investigation of glycoprotein profiles in plasma from gastric cancer patients versus healthy volunteers and to identify glycoprotein biomarkers for the early clinical detection of gastric cancer. Three novel PTMs, HexHexNAc, trimethylation and FAD, in Con A-bound ITIH3 were identified and built in molecular modeling. The aspartic acid-(290) trimethylation site was located in a metal ion-dependent adhesion site (MIDAS motif;(290)-DXSXS...T...D-(313)) that may influence important function for binding protein ligands. (C) 2013 Elsevier B.V. All rights reserved.
    關聯: Journal of Proteomics, v.83 pp.197-213
    顯示於類別:[生物科技系(所)] 期刊論文


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