Abundance-Ratio-Based Semiquantitative Analysis of Site-Specific N-Linked Glycopeptides Present in the Plasma of Hepatocellular Carcinoma Patients

• DOI: 10.1021/pr4011519
• Publication Date: 2014-03-17T10:45:53Z
• Authors: Hyoung-Joo Lee; Hyun-Jeong Cha; Jong-Sun Lim; Sun Hee Lee; Si Young Song; Hoguen Kim; William S. Hancock; Jong Shin Yoo; Young-Ki Paik
• Keywords: Male; Proteomics; 0301 basic medicine; Glycomics/methods; Vitronectin/metabolism; Tandem Mass Spectrometry; Glycopeptides/metabolism*; Tumor/metabolism; HCC; Hepatocellular/blood; Glycomics; Chromatography; Liquid; Liver Neoplasms/blood; Liver Neoplasms; Glycopeptides; Middle Aged; Vitronectin/blood; haptoglobin; 3. Good health; Polysaccharides/metabolism*; Tumor/blood; Proteomics/methods; Adult; Carcinoma, Hepatocellular; Molecular Sequence Data; Liver Neoplasms/metabolism*; vitronectin; Glycopeptides/chemistry; 03 medical and health sciences; Hepatocellular/pathology; Polysaccharides; Biomarkers, Tumor; Humans; Amino Acid Sequence; Vitronectin; Aged; Neoplasm Staging; Binding Sites; Carcinoma; N-linked glycopeptides; Reproducibility of Results; Liver Neoplasms/pathology; quantification; TMT; Polysaccharides/blood; Glycopeptides/blood; Biomarkers; Hepatocellular/metabolism*; Chromatography, Liquid

Aberrant structures of site-specific N-linked glycans are closely associated with the tumorigenesis of hepatocellular carcinoma (HCC), one of the most common fatal cancers worldwide. Vitronectin (VTN) is considered a candidate glycobiomarker of HCC. In this study, we describe a reliable and simple quantification strategy based on abundance ratios of site-specific N-linked glycopeptides of VTN to screen for potential biomarkers. A total of 14 unique N-linked glycans corresponding to 27 unique N-linked glycopeptides were characterized at three N-linked sites (Asn-86, -169, and -242) present in VTN. These glycans could be good candidate markers for HCC. Among these glycans, the abundance ratio of two representative glycoforms (fucosyl vs non-fucosyl) was significantly increased in HCC plasma relative to normal plasma. This strategy was also successfully applied to another potential HCC biomarker, haptoglobin. Furthermore, we demonstrate that our approach employing tandem mass tag (TMT) and target N-linked glycopeptides of VTN is a useful tool for quantifying specific glycans in HCC plasma relative to normal plasma. Our strategy represents a simple and potentially useful screening platform for the discovery of cancer-specific glycobiomarkers.