Glycobiology Advance Access originally published online on May 17, 2004
Glycobiology 2004 14(9):793-804; doi:10.1093/glycob/cwh094
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycobiology vol. 14 no. 9 © Oxford University Press 2004; all rights reserved.
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 Faculty of Pharmaceutical Sciences, Kinki University, Kowakae 3-4-1, Higashi-osaka 577-8502, Japan; 3 National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba 305-8602, Japan; and 4 Faculty of Agriculture, Meiji University, Higashi-Mita 1-1-1, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
Received on March 19, 2004; revised on May 5, 2004; accepted on May 5, 2004
We developed capillary affinity electrophoresis (CAE) to analyze the molecular interaction between carbohydrate chains and proteins in solution state (Nakajima et al. [2003] J. Proteome Res., 2, 81–88). 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 triantennary oligosaccharides with Gal residues at nonreducing termini and a Fuc residue linked through 
(1-6) linkage at chitobiose portion of the reducing termini but not tetraantennary 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.
1 To whom correspondence should be addressed; e-mail: k_kakehi{at}phar.kindai.ac.jp
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Debray, B. Coddeville, L. R Bomfim, and M. V Ramos A simple micro-method for determining precise oligosaccharidic specificity of mannose-binding lectins Glycobiology, December 1, 2009; 19(12): 1417 - 1426. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Kaku, H. Kaneko, N. Minamihara, K. Iwata, E. T. Jordan, M. A. Rojo, N. Minami-Ishii, E. Minami, S. Hisajima, and N. Shibuya Elderberry Bark Lectins Evolved to Recognize Neu5Ac{alpha}2,6Gal/GalNAc Sequence from a Gal/GalNAc Binding Lectin Through the Substitution of Amino-Acid Residues Critical for the Binding to Sialic Acid J. Biochem., September 1, 2007; 142(3): 393 - 401. [Abstract] [Full Text] [PDF]
|
|