Carbohydrate-Based Probes for Detection of Cellular Lectins

Galanina, Oxana; Tuzikov, Alexander; Rapoport, E. M.; Pendu, Jacques Le; Bovin, Nicolai V.

doi:10.1006/abio.1998.2859

Cited by 28 publications

(15 citation statements)

References 14 publications

(10 reference statements)

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“…Polyacrylamide-based glycoconjugates that have predetermined properties, such as molecular mass, solubility, matrix flexibility, distance between the glycotopes, and density of substitution by the different glycotopes, are a suitable tool for measuring interactions between lectins and their carbohydrate receptors (4,9,10). The adhesion of bacteria to these neoglycoconjugates can be studied by two different methods: an adhesion assay with unlabeled glycoconjugates applied to a microtiter plate or a flow cytometry assay with polyacrylamide-based glycoconjugates labeled with fluorescein.…”

Section: Discussionmentioning

confidence: 99%

Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of Pseudomonas aeruginosa

et al. 2001

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Section: Discussionmentioning

confidence: 99%

Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of Pseudomonas aeruginosa

et al. 2001

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“…Nonylphenyl Polyethylene Glycol (NP40) was purchased from Calbiochem EMD Millipore (Billerica, MA, USA). Fluoresceinated and biotinylated glycan probes (Galanina et al, 1998) were from Lectinity (Moscow, Russia).…”

Section: Methodsmentioning

confidence: 99%

Multiple proteins from ejaculated and epididymal porcine spermatozoa bind glycan motifs found in the oviduct

et al. 2014

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SUMMARYPreviously, we demonstrated that porcine spermatozoa bind selectively to Lewis x (Le X ) and bi-antennary a2-6 sialylated N-acetyllactosamine (bi-SiaLN) glycan motifs found in the oviduct. Recognition of these motifs mediates sperm binding to oviduct epithelial cells to form a reservoir. Here, we characterize proteins that bind to Le X and bi-SiaLN in ejaculated and cauda epididymal spermato-) and bi-SiaLN were used to locate the binding proteins on spermatozoa. Binding to N-acetyllactosamine (LN), a glycan that spermatozoa bind less, was assessed as a control. Over 60% of ejaculated and epididymal spermatozoa bound to suLe X and bi-SiaLN, but only 25% bound to LN. Specific binding to suLe X and bi-SiaLN required divalent cations. Fluoresceinated suLe X bound mostly to the plasma membrane at the apex of the head, whereas bi-SiaLN bound to a broader area of the head. Further characterization of receptors was accomplished by probing, with specific glycans, sperm proteins that had been separated by SDS-PAGE and transferred to blots. Glycan blot assays identified several suLe X -binding bands that migrated as 12-250 kDa. Migration of some of the bands that bound bi-SiaLN was similar to those that bound suLe X . A 12-kDa band protein was detected in seminal fluid and ejaculated spermatozoa, but absent in epididymal spermatozoa. In summary, we determined that: (i) multiple glycan-binding proteins recognize suLe X and bi-SiaLN motifs, (ii) specific binding required divalent cations and (iii) glycanbinding proteins were identified before and after sperm contact with accessory gland fluids. While the identity of these molecules is still unknown, this information predicts characteristics of the authentic oviduct suLe X and bi-SiaLN receptors on spermatozoa.

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“…In addition, 50 µg/ml zeocin or 50 µg/ml G418 were added to CHO-E and CHO-CTLA4-Ig transfectants, respectively. CHO-E was generated by transfection of CHO cells with a plasmid containing human E-selectin as described [24]. Murine fibroblast cell line, DAP-3, was cultured in DMEM with 10% FCS, 2 mM L-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin.…”

Section: Cell Lines and Primary Endothelial Cells Hsvec And Vsmcmentioning

confidence: 99%

Antibody targeted gene transfer to endothelium

Tan

Manunta

Ardjomand

et al. 2002

The Journal of Gene Medicine

106

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We have shown that our novel vector is capable of in vitro and ex vivo gene delivery to cells and human tissues including cornea, artery and vein. In particular, an Ab against E-selectin was effective at selectively delivering genes to activated endothelial cells expressing the adhesion molecule. Such a strategy will have applications for targeting these tissues prior to transplantation or autologous grafting, and, in the longer term, may allow in vivo targeting of gene therapy to inflammatory sites.

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Carbohydrate-Based Probes for Detection of Cellular Lectins

Cited by 28 publications

References 14 publications

Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of Pseudomonas aeruginosa

Recognition of Lewis x Derivatives Present on Mucins by Flagellar Components of Pseudomonas aeruginosa

Multiple proteins from ejaculated and epididymal porcine spermatozoa bind glycan motifs found in the oviduct

Antibody targeted gene transfer to endothelium

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