Quantum-dye labeled proteins for glycobiology: a viable nonradioactive alternative tracer.
Quantum dye (QD), a macrocyclic europium-chelate, developed as a cytological marker, has never been used for quantitative applications. It would be ideal, however, if the same tracer can be used for both qualitative and quantitative purposes. We have labeled some lectins and neoglycoproteins with QD for the purpose of quantitative analyses in glycobiology, and tested its suitability in three different areas in glycobiology: (1) glycosyltransferase, (2) an animal lectin - mannose-binding protein, and (3) the Gal/GalNAc receptor of rat liver membrane. Usefulness of QD-labeled lectins was amply demonstrated by the quantification of galactosyltransferase activity using QD-soybean agglutinin and QD-RCA120 ( Ricinus communis agglutinin). We also showed that QD-labeled neoglycoproteins, QD-Man-BSA and QD-Gal-BSA, can replace radioiodinated counterparts in the binding assays of animal lectins (serum mannose binding protein and hepatic Gal/GalNAc receptor.) The advantage of QD and other europium labels is that it does not decay as radioiodides do. The long shelf-life results in more consistent results from repeated experiments.
Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.
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YC
Lee
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Kawasaki
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Lee
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Suzuki
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D005063:Europium / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D005700:Galactosyltransferases / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry*; D006207:Half-Life; D007202:Indicators and Reagents / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry; D037601:Mannose-Binding Lectin; D009942:Organometallic Compounds / Q000737:chemistry*; D017785:Photons; D051381:Rats
D002352:Carrier Proteins; D005456:Fluorescent Dyes; D006023:Glycoproteins; D007202:Indicators and Reagents; D037102:Lectins; D037601:Mannose-Binding Lectin; D009942:Organometallic Compounds; C114158:quantum dye macrocyclic europium-chelate; C069858:mannose binding protein A; D005063:Europium; D005700:Galactosyltransferases