title: Identification of the binding roles of terminal and internal glycan epitopes using enzymatically synthesized N-glycans containing tandem epitopes.
abstract: Glycans play diverse roles in a wide range of biological processes. Research on glycan-binding events is essential for learning their biological and pathological functions. However, the functions of terminal and internal glycan epitopes exhibited during binding with glycan-binding proteins (GBPs) and/or viruses need to be further identified. Therefore, a focused library of 36 biantennary asparagine (Asn)-linked glycans with some presenting tandem glycan epitopes was synthesized via a combined Core Isolation/Enzymatic Extension (CIEE) and one-pot multienzyme (OPME) synthetic strategy. These N-glycans include those containing a terminal sialyl N-acetyllactosamine (LacNAc), sialyl Lewis x (sLex) and Siaα2-8-Siaα2-3/6-R structures with N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc) sialic acid form, LacNAc, Lewis x (Lex), α-Gal, and Galα1-3-Lex; and tandem epitopes including α-Gal, Lex, Galα1-3-Lex, LacNAc, and sialyl LacNAc, presented with an internal sialyl LacNAc or 1-2 repeats of an internal LacNAc or Lexcomponent. They were synthesized in milligram-scale, purified to over 98% purity, and used to prepare a glycan microarray. Binding studies using selected plant lectins, antibodies, and viruses demonstrated, for the first time, that when interpreting the binding between glycans and GBPs/viruses, not only the structure of the terminal glycan epitopes, but also the internal epitopes and/or modifications of terminal epitopes needs to be taken into account.
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Yunpeng
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Cheng
authors.initials: C
authors.lastname: Ma
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.affiliation: College of Life Science, Hebei Normal University, Shijiazhuang, Hebei 050024, China. guanwanyi@mail.hebtu.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Bai
authors.affiliation: Departments of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Byrd-Leotis
authors.affiliation: Department of Biochemistry and Emory Comprehensive Glycomics Core, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lasanajak
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Yuxi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Liuqing
authors.initials: L
authors.lastname: Wen
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: He
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Jing
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Chemistry, University of California, Davis, CA 95616, USA. xiichen@ucdavis.edu.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Li
authors.affiliation: Departments of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Steinhauer
authors.affiliation: Department of Biochemistry and Emory Comprehensive Glycomics Core, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: David F
authors.initials: DF
authors.lastname: Smith
authors.affiliation: College of Life Science, Hebei Normal University, Shijiazhuang, Hebei 050024, China. guanwanyi@mail.hebtu.edu.cn.
authors.forename: Baohua
authors.initials: B
authors.lastname: Zhao
authors.affiliation: Department of Chemistry, University of California, Davis, CA 95616, USA. xiichen@ucdavis.edu.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu and College of Life Science, Hebei Normal University, Shijiazhuang, Hebei 050024, China. guanwanyi@mail.hebtu.edu.cn.
authors.forename: Wanyi
authors.initials: W
authors.lastname: Guan
authors.affiliation: Department of Chemistry and Center of Diagnostics & Therapeutics, Georgia State University, Atlanta, GA 30303, USA. pwang11@gsu.edu.
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
mesh_terms: D060326:Chemistry Techniques, Synthetic; D004798:Enzymes / Q000378:metabolism*; D000939:Epitopes / Q000737:chemistry* / Q000378:metabolism*; D011134:Polysaccharides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004798:Enzymes; D000939:Epitopes; D011134:Polysaccharides