title: Investigation on glycosylation patterns of proteins from human liver cancer cell lines based on the multiplexed proteomics technology.
abstract: Glycosylation, a very important post-translational modification of proteins, is increasingly coming into notice. However, large-scale, throughput investigations on glycosylated proteins are few. We applied a sensitive and fast fluorescence-based multiplexed proteomics (MP) technology which included two-dimensional gel electrophoresis (2-DE) followed by the fluorescence staining of glycoprotein and mass spectrometry identification for the purpose of constructing glycoprotein databases of the typical human hepatocellular carcinoma cell lines including Hep3B cell line without metastasis and MHCC97H with highly metastatic potential as well as the control non-tumor Chang liver cell. 74+/-2 (n=3), 78+/-3 (n=3) and 72+/-5 (n=3) glycoprotein spots were detected on 2-DE gels from Chang liver, Hep3B and MHCC97H cell sample using this MP technique, respectively. In all, 80 glycoproteins from three cell lines were successfully identified via peptide mass profiling using MALDI-TOF-MS/MS and the identified glycoproteins were annotated to our databases. In addition, we also found the glycosylation pattern differences among these three cell lines. The protein glycosylation alteration would be have great significance for the diagnosis of HCC and prediction of its metastasis. This study described the construction of glycosylation patterns of proteins and glycoproteome databases of human liver cells by the novel technological platform. The glycoproteome databases also provide essential basis for following study.
authors.affiliation: Liver Cancer Institute, Zhongshan Hospital, and Department of Chemistry, Fudan University, Shanghai 200032, China.
authors.forename: Haijun
authors.initials: H
authors.lastname: Zhou
authors.forename: Yinkun
authors.initials: Y
authors.lastname: Liu
authors.forename: Jiefeng
authors.initials: J
authors.lastname: Chui
authors.forename: Kun
authors.initials: K
authors.lastname: Guo
authors.forename: Qiangling
authors.initials: Q
authors.lastname: Shun
authors.forename: Wenjing
authors.initials: W
authors.lastname: Lu
authors.forename: Hong
authors.initials: H
authors.lastname: Jin
authors.forename: Liming
authors.initials: L
authors.lastname: Wei
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D010449:Peptide Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods*
chemical_list: D006023:Glycoproteins