Glycoprotein Labeling and Detection: Novel Click Chemistry-based Applications for gel Electrophoresis, Flow Cytometry and Fluorescence Microscopy
Abstract
We demonstrate highly-selective and sensitive labeling methods for the detection of specific glycoprotein subclasses, including cell surface N- and O-linked glycoproteins and intracellular O-GlcNAc modified proteins, utilizing the copper-catalyzed cycloaddition reaction between azides and alkynes, or click chemistry. The two-step labeling technique involves the incorporation of unnatural azide-modified sugars into protein glycan structures and subsequent ligation with fluorescent or UV-excitable azide-reactive detection probes.
We demonstrate highly-selective and sensitive labeling methods for the detection of specific glycoprotein subclasses, including cell surface N- and O-linked glycoproteins and intracellular O-GlcNAc modified proteins, utilizing the copper-catalyzed cycloaddition reaction between azides and alkynes, or click chemistry. The two-step labeling technique involves the incorporation of unnatural azide-modified sugars into protein glycan structures and subsequent ligation with fluorescent or UV-excitable azide-reactive detection probes.
