Unprecedented Affinity Labeling of Carbohydrate-Binding Proteins with s-Triazinyl Glycosides

Abstract

Carbohydrate−protein interactions trigger a wide range of biological signaling pathways, the mainstays of physiological and pathological processes. However, there are an incredible number of carbohydrate-binding proteins (CBPs) that remain to be identified and characterized. This study reports for the first time the covalent labeling of CBPs by triazinyl glycosides, a new and promising class of affinitybased glycoprobes. Mono-and bis-clickable triazinyl glycosides were efficiently synthesized from unprotected oligosaccharides (chitinpentaose and 2′-fucosyl-lactose) in a single step. These molecules allow the specific covalent labeling of chitin-oligosaccharide-binding proteins (wheat germ agglutinin WGA and Bc ChiA1 D202A, an inactivated chitinase) and fucosyl-binding lectin (UEA-I), respectively.

Evolution of Lipochitooligosaccharide Binding to a LysM-RLK for Nodulation in Medicago truncatula

“…The CO5-biot was derived from the CO5-propargyl (Masselin et al 2019). CO5-propargyl (5 mg, 4 µmol) and azide-Peg3-biotin conjugate (Aldrich) (2.7 mg, 6 µmol) were mixed in 0.1 M phosphate buffer (600 µL, pH 7.4), and copper sulfate (100 µL of an aqueous solution at 10 mg/mL, 4 µmol) and sodium ascorbate (100 µL of an aqueous solution at 10 mg/mL, 5 µmol) were successively added.…”