N-glycoprotein enrichment by lectin affinity chromatography.

ABSTRACT

Lectins are proteins that bind to sugars with varying specificities and several have been identified that show differential binding to structurally variable glycans attached to glycoproteins. Consequently, lectin affinity chromatography represents a valuable tool for glycoproteome studies, allowing enrichment of glycoproteins in samples prior to their identification by mass spectrometry (MS). From the perspective of plant scientists, lectin enrichment has proven useful for studies of the proteomes of the secretory pathways and cell wall, due to the high proportion of constituent proteins that are glycosylated. This chapter outlines a strategy to generate samples enriched with glycoproteins from bulk plant tissues prior to further characterization by MS, or other techniques.