title: Glycoprobes as a tool for the study of lectins expressed on tumor cells.
abstract: Polyacrylamide glycoconjugates, Glyc-PAA, having various tags or labels are convenient tools for analysis of cellular lectins. Adaptation of such glycoprobes for flow cytometry allows us to reveal lectins expressed on cell surface and analyze their carbohydrate specificity as well as functionality. Localization of lectins is visualized by labeling of cells with fluorescein-tagged glycoprobes, Glyc-PAA-fluo, in combination with fluorescent microscopy techniques. Additionally, biotinylated glycoprobes can be immobilized on magnetic particles making it possible to separate a cell population according to its carbohydrate-binding profile. Here, we exemplify application of glycoprobes in the study of cellular siglecs and galectins, as well as lectin patterning of tumor cells. The specificity of sialic acid binding membrane-anchored lectins, siglecs-1, -5, -7, -8 and -9 was determined using this methodology. To study the carbohydrate-binding profile of soluble galactoside-binding lectins, galectins-1 or -3, these were loaded on (initially galectin free) Raji cells and probed using Glyc-PAA-fluo. Lessons learned from this model system allowed us to study the galectin distribution pattern of tumors: cells obtained from mice carrying mammary adenocarcinoma or lymphoma were probed with Glyc-PAA-fluo using flow cytometry. Disaccharide 6OSuLacdiNAc was shown to be the most potent probe for adenocarcinoma cells, demonstrating that 6OSuLacdiNAc-binding molecules accumulate on cell surface in a patch-wise distribution.
authors.affiliation: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, 117997, Moscow, Russia.
authors.forename: Olga
authors.initials: O
authors.lastname: Kurmyshkina
authors.forename: Eugenia
authors.initials: E
authors.lastname: Rapoport
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Moiseeva
authors.forename: Elena
authors.initials: E
authors.lastname: Korchagina
authors.forename: Tatiana
authors.initials: T
authors.lastname: Ovchinnikova
authors.forename: Galina
authors.initials: G
authors.lastname: Pazynina
authors.forename: Ivan
authors.initials: I
authors.lastname: Belyanchikov
authors.forename: Nicolai
authors.initials: N
authors.lastname: Bovin
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D019793:Fluorescein / Q000737:chemistry; D005697:Galactosides / Q000737:chemistry / Q000378:metabolism; D037161:Galectins / Q000378:metabolism*; D006801:Humans; D008223:Lymphoma / Q000378:metabolism / Q000473:pathology*; D008280:Magnetics; D008297:Male; D015674:Mammary Neoplasms, Animal / Q000378:metabolism / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D015335:Molecular Probes / Q000737:chemistry*; D011485:Protein Binding; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D019793:Fluorescein; D005697:Galactosides; D037161:Galectins; D008562:Membrane Glycoproteins; D015335:Molecular Probes; D000099166:Polyacrylamides