title: Cross-comparison of protein recognition of sialic acid diversity on two novel sialoglycan microarrays.
abstract: DNA and protein arrays are commonly accepted as powerful exploratory tools in research. This has mainly been achieved by the establishment of proper guidelines for quality control, allowing cross-comparison between different array platforms. As a natural extension, glycan microarrays were subsequently developed, and recent advances using such arrays have greatly enhanced our understanding of protein-glycan recognition in nature. However, although it is assumed that biologically significant protein-glycan binding is robustly detected by glycan microarrays, there are wide variations in the methods used to produce, present, couple, and detect glycans, and systematic cross-comparisons are lacking. We address these issues by comparing two arrays that together represent the marked diversity of sialic acid modifications, linkages, and underlying glycans in nature, including some identical motifs. We compare and contrast binding interactions with various known and novel plant, vertebrate, and viral sialic acid-recognizing proteins and present a technical advance for assessing specificity using mild periodate oxidation of the sialic acid chain. These data demonstrate both the diversity of sialic acids and the analytical power of glycan arrays, showing that different presentations in different formats provide useful and complementary interpretations of glycan-binding protein specificity. They also highlight important challenges and questions for the future of glycan array technology and suggest that glycan arrays with similar glycan structures cannot be simply assumed to give similar results.
authors.affiliation: Department of Medicine and Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Vered
authors.initials: V
authors.lastname: Padler-Karavani
authors.forename: Xuezheng
authors.initials: X
authors.lastname: Song
authors.forename: Hai
authors.initials: H
authors.lastname: Yu
authors.forename: Nancy
authors.initials: N
authors.lastname: Hurtado-Ziola
authors.forename: Shengshu
authors.initials: S
authors.lastname: Huang
authors.forename: Saddam
authors.initials: S
authors.lastname: Muthana
authors.forename: Harshal A
authors.initials: HA
authors.lastname: Chokhawala
authors.forename: Jiansong
authors.initials: J
authors.lastname: Cheng
authors.forename: Andrea
authors.initials: A
authors.lastname: Verhagen
authors.forename: Martijn A
authors.initials: MA
authors.lastname: Langereis
authors.forename: Ralf
authors.initials: R
authors.lastname: Kleene
authors.forename: Melitta
authors.initials: M
authors.lastname: Schachner
authors.forename: Raoul J
authors.initials: RJ
authors.lastname: de Groot
authors.forename: Yi
authors.initials: Y
authors.lastname: Lasanajak
authors.forename: Haruo
authors.initials: H
authors.lastname: Matsuda
authors.forename: Richard
authors.initials: R
authors.lastname: Schwab
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.forename: David F
authors.initials: DF
authors.lastname: Smith
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
authors.forename: Ajit
authors.initials: A
authors.lastname: Varki
mesh_terms: D000107:Acetylation; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D015703:Antigens, CD / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D006017:Glycolipids / Q000276:immunology / Q000378:metabolism*; D054794:Glycomics / Q000295:instrumentation / Q000379:methods / Q000592:standards; D037102:Lectins / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000276:immunology / Q000378:metabolism*; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000378:metabolism; D037121:Plant Lectins / Q000378:metabolism; D011134:Polysaccharides / Q000276:immunology / Q000378:metabolism*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods / Q000592:standards; D015203:Reproducibility of Results; D063265:Sialic Acid Binding Immunoglobulin-like Lectins
chemical_list: D000906:Antibodies; D015703:Antigens, CD; C490650:CHL1 protein, human; D015815:Cell Adhesion Molecules; D006017:Glycolipids; D037102:Lectins; D037121:Plant Lectins; D011134:Polysaccharides; C412227:SIGLEC9 protein, human; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; C009255:sialoglycolipids; D010504:Periodic Acid; C009288:metaperiodate; D019158:N-Acetylneuraminic Acid