Glycobiology Advance Access published online on May 17, 2004
Glycobiology, doi:10.1093/glycob/cwh094
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1 Faculty of Pharmaceutical Sciences, Kinki University, Kowakae 3-4-1, Higashi-osaka 577-8502, Japan
* To whom correspondence should be addressed. E-mail: k_kakehi{at}phar.kindai.ac.jp.
We developed capillary affinity electrophoresis (CAE) to analyze the molecular interaction between carbohydrate chains and proteins in solution state (Nakajima et al., 2003). A mixture of oligosaccharides derived from a glycoprotein was labeled with 8-aminopyrene-1,3,6-trisulfonate (APTS), and used as glycan library without isolation. Interaction of a carbohydrate-binding protein with each oligosaccharide in the mixture could be simultaneously observed, and relative affinities of oligosaccharides toward the protein were accurately determined. In this study, we applied CAE to detect the presence of lectins in some plants (Japanese elderberry bark and tulip bulb). In the crude extract of the elderberry bark, binding activity toward sialo-carbohydrate chains could be easily detected. We also examined the presence of lectins in the crude extract of tulip bulbs and determined the detailed carbohydrate-binding specificity of Tulipa gesneriana agglutinin (TGA), one of the lectins from tulip bulbs. Kinetic studies demonstrated that TGA showed novel carbohydrate-binding specificity and preferentially recognized tri-antennary oligosaccharides having Gal residues at non-reducing termini and a Fuc residue linked through
Revised May 5, 2004
Accepted May 5, 2004
ORIGINAL ARTICLES
Screening method of carbohydrate-binding proteins in biological sources by capillary affinity electrophoresis and its application to determination of Tulipa gesneriana agglutinin in tulip bulbs
2 National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba 305-8602, Japan
3 Faculty of Agriculture, Meiji University, Higashi-Mita 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
Abstract 
(1-6)linkage at chitobiose portion of the reducing termini, but not tetra-antennary carbohydrates. The results described here indicate that CAE will be a valuable method for both screening of lectins in natural sources and determination of their detailed carbohydrate-binding specificities.
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