Top-Down Chemoenzymatic Approach to Synthesizing Diverse High-Mannose N-Glycans and Related Neoglycoproteins for Carbohydrate Microarray Analysis

Christian Toonstra, Lisa Wu, Chao Li, Denong Wang, Lai Xi Wang

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

High-mannose-type N-glycans are an important component of neutralizing epitopes on HIV-1 envelope glycoprotein gp120. They also serve as signals for protein folding, trafficking, and degradation in protein quality control. A number of lectins and antibodies recognize high-mannose-type N-glycans, and glycan array technology has provided an avenue to probe these oligomannose-specific proteins. We describe in this paper a top-down chemoenzymatic approach to synthesize a library of high-mannose N-glycans and related neoglycoproteins for glycan microarray analysis. The method involves the sequential enzymatic trimming of two readily available natural N-glycans, the Man 9 GlcNAc 2 Asn prepared from soybean flour and the sialoglycopeptide (SGP) isolated from chicken egg yolks, coupled with chromatographic separation to obtain a collection of a full range of natural high-mannose N-glycans. The Asn-linked N-glycans were conjugated to bovine serum albumin (BSA) to provide neoglycoproteins containing the oligomannose moieties. The glycoepitopes displayed were characterized using an array of glycan-binding proteins, including the broadly virus-neutralizing agents, glycan-specific antibody 2G12, Galanthus nivalis lectin (GNA), and Narcissus pseudonarcissus lectin (NPA).

Original languageEnglish (US)
Pages (from-to)1911-1921
Number of pages11
JournalBioconjugate Chemistry
Volume29
Issue number6
DOIs
StatePublished - Jun 20 2018

Bibliographical note

Funding Information:
This work was supported in part by NIH grants R01AI113896 (to L.X.W.), R56AI118464 (to D.W.), and R21AI124068 (to D.W.).

Publisher Copyright:
© 2018 American Chemical Society.

Fingerprint

Dive into the research topics of 'Top-Down Chemoenzymatic Approach to Synthesizing Diverse High-Mannose N-Glycans and Related Neoglycoproteins for Carbohydrate Microarray Analysis'. Together they form a unique fingerprint.

Cite this