title: BAD-lectins: boronic acid-decorated lectins with enhanced binding affinity for the selective enrichment of glycoproteins.
abstract: The weak and variable binding affinities exhibited by lectin-carbohydrate interactions have often compromised the practical utility of lectin in capturing glycoproteins for glycoproteomic applications. We report here the development and applications of a new type of hybrid biomaterial, namely a boronic acid-decorated lectin (BAD-lectin), for efficient bifunctional glycoprotein labeling and enrichment. Our binding studies showed an enhanced affinity by BAD-lectin, likely to be mediated via the formation of boronate ester linkages between the lectin and glycan subsequent to the initial recognition process and thus preserving its glycan-specificity. Moreover, when attached to magnetic nanoparticles (BAD-lectin@MNPs), 2 to 60-fold improvement on detection sensitivity and enrichment efficiency for specific glycoproteins was observed over the independent use of either lectin or BA. Tested at the level of whole cell lysates for glycoproteomic applications, three different types of BAD-lectin@MNPs exhibited excellent specificities with only 6% overlapping among the 295 N-linked glycopeptides identified. As many as 236 N-linked glycopeptides (80%) were uniquely identified by one of the BAD-lectin@MNPs. These results indicated that the enhanced glycan-selective recognition and binding affinity of BAD-lectin@MNPs will facilitate a complementary identification of the under-explored glycoproteome.
authors.affiliation: Department of Chemistry, National Tsing Hua University, Hsinchu 300-71, Taiwan.
authors.forename: Ying-Wei
authors.initials: YW
authors.lastname: Lu
authors.forename: Chih-Wei
authors.initials: CW
authors.lastname: Chien
authors.forename: Po-Chiao
authors.initials: PC
authors.lastname: Lin
authors.forename: Li-De
authors.initials: LD
authors.lastname: Huang
authors.forename: Chang-Yang
authors.initials: CY
authors.lastname: Chen
authors.forename: Sz-Wei
authors.initials: SW
authors.lastname: Wu
authors.forename: Chia-Li
authors.initials: CL
authors.lastname: Han
authors.forename: Kay-Hooi
authors.initials: KH
authors.lastname: Khoo
authors.forename: Chun-Cheng
authors.initials: CC
authors.lastname: Lin
authors.forename: Yu-Ju
authors.initials: YJ
authors.lastname: Chen
mesh_terms: D000818:Animals; D001897:Boronic Acids / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D006023:Glycoproteins / Q000032:analysis* / Q000378:metabolism*; D006367:HeLa Cells; D006736:Horses; D006801:Humans; D037102:Lectins / Q000032:analysis* / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D001897:Boronic Acids; D006023:Glycoproteins; D037102:Lectins