Mucin-lectin interactions assessed by flow cytometry.
The O-glycosylated domains of mucins and mucin-type glycoproteins contain 50-80% of carbohydrate and possess expanded conformations. Herein, we describe a flow cytometry (FCM) method for determining the carbohydrate-binding specificities of lectins to mucin. Biotinylated mucin was immobilized on streptavidin-coated beads, and the binding specificities of the major mucin sugar chains, as determined by GC-MS and MALDI-ToF, were monitored using fluorescein-labeled lectins. The specificities of lectins toward specific biotinylated glycans were determined as controls. The advantage of flexibility, multiparametric data acquisition, speed, sensitivity, and high-throughput capability makes flow cytometry a valuable tool to study diverse interactions between glycans and proteins.
Jeffers
Faye
F
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK.
Fuell
Christine
C
Tailford
Louise E
LE
Mackenzie
Donald A
DA
Bongaerts
Roy J
RJ
Juge
Nathalie
N
D000818:Animals; D005434:Flow Cytometry; D009077:Mucins / Q000378:metabolism*; D037121:Plant Lectins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding
D009077:Mucins; D037121:Plant Lectins; D011134:Polysaccharides