Detection and Isolation of Lectin-Transfected COS Cells Based on Cell Adhesion to Immobilized Glycosphingolipids

Yang, Lynda J.‐S.; Zeller, Cynthia B.; Schnaar, Ronald L.

doi:10.1006/abio.1996.0145

Cited by 19 publications

(13 citation statements)

References 3 publications

(3 reference statements)

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“…Microplate Cell Adhesion to Adsorbed Glycolipids-Adhesion was performed as reported previously (18,35). Briefly, a 50-l aliquot of ethanol/water (1:1) containing gangliosides (concentrations as indicated), phosphatidylcholine (0.5 M), and cholesterol (2.0 M) was added to microwells (96-well Serocluster; Costar, Cambridge, MA).…”

Section: Methodsmentioning

confidence: 99%

“…Treatment of GD1a with iodoethane converted its sialic acids to their ethyl esters (5), which were then converted to their 1-amides (6) or reduced to the corresponding 1-alcohols (7). mersed upright in a vat of PBS, inverted, and placed in an immersed custom-designed Plexiglas box, which was sealed with a gasket to exclude air (35). The inverted plate in its fluid-filled chamber was placed in a centrifuge carrier and centrifuged at 110 ϫ g to gently remove nonadherent cells.…”

Section: Methodsmentioning

confidence: 99%

“…The decrease in absorbance at 340 nm as a function of time was measured simultaneously in each well using a Molecular Devices UV multiwell kinetic plate reader. Details of the adhesion assay have been described (35).…”

Section: Methodsmentioning

confidence: 99%

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Sialic Acid Specificity of Myelin-associated Glycoprotein Binding

Collins

Yang

Mukhopadhyay

et al. 1997

Journal of Biological Chemistry

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165

108

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Sialic Acid Specificity of Myelin-associated Glycoprotein Binding

Collins

Yang

Mukhopadhyay

et al. 1997

Journal of Biological Chemistry

Self Cite

165

108

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show abstract

“…Microplate Cell Adhesion to Adsorbed Glycolipids-Adhesion was performed as reported previously (20,22,27). Aliquots (50 l) of ethanol/water (1:1) containing phosphatidylcholine (0.5 M), cholesterol (2.0 M), and gangliosides (concentrations as indicated) were added to microwells (96-well Serocluster, Costar Corp., Cambridge, MA).…”

Section: Methodsmentioning

confidence: 99%

“…To gently remove nonadherent cells after the incubations, plates were immersed in phosphatebuffered saline, inverted, and placed in an immersed Plexiglas box that was sealed with a gasket to exclude air (27). The inverted plate in its fluid-filled chamber was placed in a centrifuge carrier and centrifuged at 110 ϫ g. The box was again immersed in phosphate-buffered saline; the plate was removed and righted (while immersed); and excess surface buffer was removed by aspiration, leaving 300 l/well.…”

Section: Methodsmentioning

confidence: 99%

Binding Specificities of the Sialoadhesin Family of I-type Lectins

Collins¹,

Kiso²,

Hasegawa³

et al. 1997

Journal of Biological Chemistry

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142

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The carbohydrate binding specificities of three sialoadhesins, a subgroup of I-type lectins (immunoglobulin superfamily lectins), were compared by measuring lectin-transfected COS cell adhesion to natural and synthetic gangliosides. The neural sialoadhesins, myelinassociated glycoprotein (MAG) and Schwann cell myelin protein (SMP), had similar and stringent binding specificities. Each required an ␣2,3-linked sialic acid on the terminal galactose of a neutral saccharide core, and they shared the following rank-order potency of binding: G Q1b␣ > > G D1a ‫؍‬ G T1b > > G M3 ‫؍‬ G M4 > > G M1 , G D1b , G D3 , G Q1b (nonbinders). In contrast, sialoadhesin had less exacting specificity, binding to gangliosides that bear either terminal ␣2,3-or ␣2,8-linked sialic acids with the following rank-order potency of binding: Sialoadhesins (1) are a structurally and functionally related family consisting of five immunoglobulin superfamily lectins (I-type lectins) (2) including myelin-associated glycoprotein (MAG), 1 Schwann cell myelin protein (SMP), CD22, CD33, and sialoadhesin. MAG and SMP are found on oligodendroglia and Schwann cells in the nervous system (3, 4), CD22 is expressed on a subset of B lymphocytes, sialoadhesin on a subset of macrophages, and CD33 on cells of myelomonocytic lineage. Sialoadhesins have been proposed to mediate cell-cell recognition, perhaps via their carbohydrate binding activities (5-7). Each sialoadhesin family member has two or more Ig-like domains: an amino-terminal V-set domain followed by one or more (up to 16) C2-set domains (8). Domain deletion and sitedirected mutagenesis of sialoadhesin and CD22 localize their carbohydrate-binding sites to the amino-terminal V-set domain, with contributions (for CD22) from the adjoining C2-set domain. These first two domains share very high amino acid sequence similarity between MAG and SMP (Ͼ70%) and significant similarity across all I-type lectins (Ͼ30% in pairwise comparisons) (2,8,9).Each I-type lectin binds to carbohydrate structures bearing a nonreducing terminal sialic acid (1,6,10). Sialic acids are a common nonreducing terminus of vertebrate glycoconjugates and appear to play uniquely important roles in recognition phenomena. Because sialic acids may be linked to Gal, GalNAc, or other sialic acid residues at various positions and because they may carry different substituents on their 9-carbon base structure, the sialic acids represent a diverse family of carbohydrate determinants (11). In certain sialic acid-dependent recognition systems, determinant stringency is low. For example, selectins bind to oligosaccharides bearing truncated sialic acids (12) or appropriately placed anionic groups (sulfates, carboxylic acids) otherwise unrelated to the sialic acid structure (13-16). In contrast, sialoadhesins appear to have more stringent sialic acid specificities (see "Discussion") (9). In this study, we used cells expressing different sialoadhesins to explore and compare the fine structural preferences of their binding to target sialylated glycoconju...

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Biological Activities of Proteins and Marine‐derived Peptides from Byproducts and Seaweeds

Hayes

2013

Marine Proteins and Peptides

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Detection and Isolation of Lectin-Transfected COS Cells Based on Cell Adhesion to Immobilized Glycosphingolipids

Cited by 19 publications

References 3 publications

Sialic Acid Specificity of Myelin-associated Glycoprotein Binding

Sialic Acid Specificity of Myelin-associated Glycoprotein Binding

Binding Specificities of the Sialoadhesin Family of I-type Lectins

Biological Activities of Proteins and Marine‐derived Peptides from Byproducts and Seaweeds

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