Detection of the Neu5 Ac (alpha 2,3) Gal (beta 1,4) GlcNAc sequence with the leukoagglutinin from Maackia amurensis: light and electron microscopic demonstration of differential tissue expression of terminal sialic acid in alpha 2,3- and alpha 2,6-linkageT Sata, PM Lackie, DJ Taatjes, W Peumans and J Roth Interdepartmental Electron Microscopy, University of Basal, Switzerland. The Maackia amurensis leukoagglutinin has been shown to react specifically with the Neu5Ac (alpha 2,3) Gal sequence of asparagine- linked complex type oligosaccharides. We report here the preparation of Maackia amurensis lectin-gold complexes and their application for light and electron microscopic detection of the Neu5 Ac (alpha 2,3) Gal sequence in various tissues. The use of the lectin directly gold labeled was superior to a two-step cytochemical affinity technique using a fetuin-gold complex. The Maackia amurensis lectin-gold staining was inhibited by pre-incubation of the lectin-gold complexes with 50 mM alpha 2,3 sialyllactose, whereas alpha 2,6 sialyllactose up to concentrations of 1 M had no effect, thus demonstrating the high specificity of the histochemical staining. In addition to N-glycanase- sensitive asparagine-linked oligosaccharides, beta-elimination- sensitive serine/threonine-linked oligosaccharides could be detected. Data are presented which show that cellular staining patterns obtained with Maackia amurensis lectin-gold complexes may differ from those with elderberry bark lectin-gold, which detects the Neu5 Ac (alpha 2,6) Gal/GalN Ac sequence. Electron microscopic double labeling for direct study of the differential distribution of the Neu5 Ac (alpha 2,3) Gal and Neu5 Ac (alpha 2,6) Gal sequences is reported. Therefore, the availability of two sialic acid binding lectins with different linkage specificity for histochemistry provides the first opportunity to study tissue and cell type expression of these terminal sequences of glycoproteins.
Volume 37,
Issue 11,
pp. 1577-1588,
11/01/1989
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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] |
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X. Bai, J. R. Brown, A. Varki, and J. D. Esko Enhanced 3-O-sulfation of galactose in Asn-linked glycans and Maackia amurenesis lectin binding in a new Chinese hamster ovary cell line Glycobiology, August 1, 2001; 11(8): 621 - 632. [Abstract] [Full Text] [PDF] |
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F. Ulloa and F. X. Real Differential Distribution of Sialic Acid in {{alpha}}2,3 and {{alpha}}2,6 Linkages in the Apical Membrane of Cultured Epithelial Cells and Tissues J. Histochem. Cytochem., April 1, 2001; 49(4): 501 - 510. [Abstract] [Full Text] |
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D. Kube, L. Adams, A. Perez, and P. B. Davis Terminal sialylation is altered in airway cells with impaired CFTR-mediated chloride transport Am J Physiol Lung Cell Mol Physiol, March 1, 2001; 280(3): L482 - L492. [Abstract] [Full Text] [PDF] |
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F. Uehara, N. Ohba, and M. Ozawa Isolation and Characterization of Mucinlike Glycoprotein Associated with Photoreceptor Cells Invest. Ophthalmol. Vis. Sci., August 1, 2000; 41(9): 2759 - 2765. [Abstract] [Full Text] |
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H. Kaku, Y. Tanaka, K. Tazaki, E. Minami, H. Mizuno, and N. Shibuya Sialylated Oligosaccharide-specific Plant Lectin from Japanese Elderberry (Sambucus sieboldiana) Bark Tissue Has a Homologous Structure to Type II Ribosome-inactivating Proteins, Ricin and Abrin J. Biol. Chem., January 19, 1996; 271(3): 1480 - 1485. [Abstract] [Full Text] [PDF] |
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