scholarly journals S18.20 Preparation of two kinds of sialic acid specific monomeric monovalent lectin derivatives and their application to the study of cell surface glycoconjugates by flow cytometry

1993 ◽  
Vol 10 (4) ◽  
pp. 333-333
Author(s):  
H. Kaku ◽  
Y. Mori ◽  
I. J. Goldstein ◽  
N. Shibuya
Keyword(s):  
Flow Cytometry ◽  
Sialic Acid ◽  
Cell Surface ◽  
Cell Surface Glycoconjugates

scholarly journals Exo-enzymatic addition of diazirine-modified sialic acid to cell surfaces enables photocrosslinking of glycoproteins

2021 ◽  
Author(s):  
Nageswari Yarravarapu ◽  
Rohit Sai Reddy Konada ◽  
Narek Darabedian ◽  
Nichole J. Pedowtiz ◽  
Soumya N. Krishnamurthy ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Cell Surface ◽  
Cell Types ◽  
Cell Surfaces ◽  
Binding Interactions ◽  
Multiple Cell ◽  
Labeling Method ◽  
Cell Surface Glycoconjugates

Glycan binding often mediates extracellular macromolecular recognition events. Accurate characterization of these binding interactions can be difficult because of dissociation and scrambling that occur during purification and analysis steps. Use of photocrosslinking methods has been pursued to covalently capture glycan-dependent interactions in situ however use of metabolic glycan engineering methods to incorporate photocrosslinking sugar analogs is limited to certain cell types. Here we report an exo-enzymatic labeling method to add a diazirine-modified sialic acid (SiaDAz) to cell surface glycoconjugates. The method involves chemoenzymatic synthesis of diazirine-modified CMP-sialic acid (CMP-SiaDAz), followed by sialyltransferase-catalyzed addition of SiaDAz to desialylated cell surfaces. Cell surface SiaDAz-ylation is compatible with multiple cell types and is facilitated by endogenous extracellular sialyltransferase activity present in Daudi B cells. This method for extracellular addition of α2-6-linked SiaDAz enables UV-induced crosslinking of CD22, demonstrating the utility for covalent capture of glycan-mediated binding interactions.

scholarly journals Trans-sialylation: a strategy used to incorporate sialic acid into oligosaccharides

2021 ◽  
Author(s):  
Rosa M. de Lederkremer ◽  
M. Eugenia Giorgi ◽  
Rosalia Agusti
Keyword(s):  
Sialic Acid ◽  
Cell Surface ◽  
Crucial Role ◽  
Synthetic Methods ◽  
Cell Surface Glycoconjugates

Sialic acid, as component of cell surface glycoconjugates, plays a crucial role in recognition events. Efficient synthetic methods are necessary for the supply of sialosides in enough quantities for biochemical...

Glycoconjugates on the surface of spores of the pathogenic fungus Phytophthora cinnamomi studied using fluorescence and electron microscopy and flow cytometry

1990 ◽  
Vol 36 (3) ◽  
pp. 183-192 ◽  
Author(s):  
A. R. Hardham ◽  
E. Suzaki
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Phytophthora Cinnamomi ◽  
Pathogenic Fungus ◽  
Fluorescein Isothiocyanate ◽  
Pathogenic Fungi ◽  
Surface Flow ◽  
Soybean Agglutinin ◽  
Binding Material

Glycoconjugates on the surface of zoospores and cysts of the pathogenic fungus Phytophthora cinnamomi have been studied using fluorescein isothiocyanate labelled lectins for fluorescence microscopy and flow cytometry, and ferritin- and gold-labelled lectins for ultrastructural analysis. Of the five lectins used, only concanavalin A (ConA) binds to the surface of the zoospores, including the flagella and water expulsion vacuole. This suggests that of accessible saccharides, glucosyl or mannosyl residues predominate on the outer surface of the zoospore plasma membrane. Early in encystment, a system of flat disc-like cisternae, which underlie the zoospore plasma membrane, vesiculate. These and other small peripheral vesicles quickly disappear. After the induction of encystment, ConA is no longer localised close to the plasma membrane but binds to material loosely associated with the cell surface. Quantitative measurements by flow cytometry indicate that the ConA-binding material is gradually lost from the cell surface. The cyst wall is weakly labelled, but the site of germ tube emergence stains intensely. During the first 2 min after the induction of encystment, material that binds soybean agglutinin, Helix pommatia agglutinin, and peanut agglutinin appears on the surface of the fungal cells. The distribution of this material, rich in galactosyl or N-acetyl-D-galactosaminosyl residues, is initially patchy, but by 5 min the material evenly coats most of the cell surface. Labelling of zoospores in which intracellular sites are accessible indicates that the soybean agglutinin binding material is stored in vesicles that lie beneath the plasma membrane. Quantitation of soybean agglutinin labelling shows that maximum binding occurs 2–3 min after the induction of encystment. Key words: cell surface, flow cytometry, lectins, pathogenic fungi, Phytophthora cinnamomi.

Metabolic expression of thiol-derivatized sialic acids on the cell surface and their quantitative estimation by flow cytometry

2006 ◽  
Vol 1 (4) ◽  
pp. 1840-1851 ◽  
Author(s):  
Srinivasa-Gopalan Sampathkumar ◽  
Mark B Jones ◽  
Kevin J Yarema
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
Quantitative Estimation ◽  
Sialic Acids

Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry

2011 ◽  
Vol 134 (2) ◽  
pp. 113-121 ◽  
Author(s):  
Laia Llinàs ◽  
Adriana Lázaro ◽  
Jose de Salort ◽  
Jessica Matesanz-Isabel ◽  
Jordi Sintes ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Surface ◽  
B Cell ◽  
Plasma Cells ◽  
Expression Profiles ◽  
Cell Surface Molecules ◽  
Surface Molecules ◽  
B Cell Subsets
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