title: Profiling of concanavalin A-binding glycoproteins in human hepatic stellate cells activated with transforming growth factor-β1.
abstract: Glycoproteins play important roles in maintaining normal cell functions depending on their glycosylations. Our previous study indicated that the abundance of glycoproteins recognized by concanavalin A (ConA) was increased in human hepatic stellate cells (HSCs) following activation by transforming growth factor-β1 (TGF-β1); however, little is known about the ConA-binding glycoproteins (CBGs) of HSCs. In this study, we employed a targeted glycoproteomics approach using lectin-magnetic particle conjugate-based liquid chromatography-tandem mass spectrometry to compare CBG profiles between LX-2 HSCs with and without activation by TGF-β1, with the aim of discovering novel CBGs and determining their possible roles in activated HSCs. A total of 54 and 77 proteins were identified in the quiescent and activated LX-2 cells, respectively. Of the proteins identified, 14.3% were glycoproteins and 73.3% were novel potential glycoproteins. Molecules involved in protein processing in the endoplasmic reticulum (e.g., calreticulin) and calcium signaling (e.g., 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase β-2 [PLCB2]) were specifically identified in activated LX-2 cells. Additionally, PLCB2 expression was upregulated in the cytoplasm of the activated LX-2 cells, as well as in the hepatocytes and sinusoidal cells of liver cirrhosis tissues. In conclusion, the results of this study may aid future investigations to find new molecular mechanisms involved in HSC activation and antifibrotic therapeutic targets.
authors.affiliation: Department of Genetics and Molecular Biology, Xi'an Jiaotong University College of Medicine, Xi'an 710061, Shaanxi, China. yannan159@mail.xjtu.edu.cn.
authors.forename: Yannan
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China. zhongyglyco@163.com.
authors.forename: Yaogang
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China. glyanglife@hotmail.com.
authors.forename: Ganglong
authors.initials: G
authors.lastname: Yang
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China. rooroo@163.com.
authors.forename: Tianran
authors.initials: T
authors.lastname: Ma
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China. csshxzza@163.com.
authors.forename: Liyuan
authors.initials: L
authors.lastname: Jia
authors.affiliation: Department of Genetics and Molecular Biology, Xi'an Jiaotong University College of Medicine, Xi'an 710061, Shaanxi, China. yannan159@mail.xjtu.edu.cn.
authors.forename: Chen
authors.initials: C
authors.lastname: Huang
authors.affiliation: Laboratory for Functional Glycomics, College of Life Sciences, Northwest University, Xi'an 710069, Shaanxi, China. zhengli@nwu.edu.cn.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
mesh_terms: D002460:Cell Line; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D063990:Gene Ontology; D055166:Hepatic Stellate Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008103:Liver Cirrhosis / Q000378:metabolism / Q000473:pathology; D060328:Magnetic Phenomena; D013058:Mass Spectrometry; D058977:Molecular Sequence Annotation; D054799:Phospholipase C beta / Q000378:metabolism; D060066:Protein Interaction Maps / Q000187:drug effects; D040901:Proteomics; D011952:Receptors, Concanavalin A / Q000302:isolation & purification / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D011952:Receptors, Concanavalin A; D053773:Transforming Growth Factor beta1; C027774:concanavalin A-binding glycoproteins; C518036:PLCB2 protein, human; D054799:Phospholipase C beta