title: Development and application of a novel recombinant Aleuria aurantia lectin with enhanced core fucose binding for identification of glycoprotein biomarkers of hepatocellular carcinoma.
abstract: The Aleuria aurantia lectin (AAL) derived from orange peel fungus contains five fucose-binding sites that recognizes fucose bound in α-1,2, α-1,3, α-1,4, and α-1,6 linkages to N-acetylglucosamine and galactose. Recently, we have created several recombinant AAL (rAAL) proteins that had altered binding affinity to fucose linkages. In this report, we further characterize the binding specificity of one of the mutated lectins, N224Q lectin. This lectin was characterized by lectin Western blotting, surface plasmon resonance, and glycan microarray and shown to have increased binding to fucosylated glycan. Subsequently, we used this lectin to identify secreted fucosylated glycoproteins from a fetal hepatic cell line. Proteomic analysis revealed several glycoproteins secreted by the fetal cell line that were bound by N224Q lectin. These findings were confirmed by subsequent proteomic analysis of human serum from control patients or patients with hepatocellular carcinoma. These represent candidate oncofetal markers for liver cancer.
authors.affiliation: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Pamela
authors.initials: P
authors.lastname: Norton
authors.affiliation: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Comunale
authors.affiliation: Graduate School of Biomedical Sciences and Professional Studies, Microbiology and Immunology Graduate Program, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Harmin
authors.initials: H
authors.lastname: Herrera
authors.affiliation: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Mengjun
authors.initials: M
authors.lastname: Wang
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Josef
authors.initials: J
authors.lastname: Houser
authors.affiliation: Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Wimmerova
authors.affiliation: Baruch S. Blumberg Institute, Doylestown, PA, USA.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Romano
authors.affiliation: Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, PA, USA.
authors.forename: Anand
authors.initials: A
authors.lastname: Mehta
mesh_terms: D001203:Ascomycota / Q000737:chemistry; D014408:Biomarkers, Tumor / Q000032:analysis / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D005643:Fucose / Q000032:analysis / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008113:Liver Neoplasms / Q000175:diagnosis / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D040901:Proteomics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D006023:Glycoproteins; D037102:Lectins; D011134:Polysaccharides; D011994:Recombinant Proteins; C064549:lectin, Aleuria aurantia; D005643:Fucose
keywords: Aleuria aurantia lectin; Biomarker; Glycoproteomics; Glycosylation; Hepatocellular carcinoma; Liver cancer