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Isolation and characterization of the potential receptor for wheat germ agglutinin from human neutrophils

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Abstract

Neutrophils participate in host protection and central to this process is the regulation of oxidative mechanisms. We purified by affinity chromatography the receptor for the GlcNAc-specific WGA from CD14+ CD16+ cell lysates (WGAr). The receptor is a 141 kDa glycoprotein constituted by two subunits of 78 and 63 kDa. It is mainly composed of Ser, Asx, and Gly, and, in a minor proportion, His, Cys, and Pro. Its glycan portion contains GlcNAc, Gal, and Man; NeuAc and GalNAc were identified in a minor proportion. The amino acid sequence of the WGA receptor was predicted from tryptic peptides by MALDI-TOF, both subunits showed homology with cytokeratin type II (26 and 29% for the 78 and 63 kDa subunits, respectively); the 78 kDa subunit showed also homology with the human transferrin receptor (24%). Antibodies against WGAr induce higher oxidative burst than WGA, determined by NBT reduction; however, this effect was inhibited (p < 0.05) with GlcNAc suggesting that WGAr participates as mediator in signal transduction in neutrophils.

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Abbreviations

BSA:

bovine serum albumin

WGA:

wheat germ agglutinin

WGAr:

wheat germ agglutinin neutrophils receptor

PMA:

phorbol 12-myristate 13-acetate

FITC:

fluorescein isothiocyanate

NBT:

nitroblue tetrazolium

GlcNAc:

N-acetyl-D-glucosamine

GalNAc:

N-acetyl-D-galactosamine

Gal:

galactose

Man:

Mannose

Fuc:

Fucose

NeuAc:

sialic acid; N-acetyl-neuraminic acid

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Authors and Affiliations

  1. Departamento de Bioquímica, Instituto Nacional de Enfermedades Respiratorias, Tlalpan, 14080, Mexico

    Carlos Solórzano

  2. Laboratoire de Chimie Biologique de la, Université des Sciences et Technologies de Lille, UMR du CNRS n^ 8576, Villeneuve d’Ascq, Cedex, 59655, France

    Stéphane Bouquelet & Marie-Christine Slomianny

  3. Unidad de Investigación, Centro Médico Nacional Siglo XXI, IMSS, 10230, Mexico

    M. Ali Pereyra & Francisco Blanco-Favela

  4. Laboratorio de Inmunología, Departamento de Bioquímica, Facultad de Medicina, UNAM, PO Box 70159, 04510 Mexico, D F, Mexico

    Raúl Chavez, Ricardo Lascurain, Edgar Zenteno & Concepción Agundis

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  1. Carlos Solórzano
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  2. Stéphane Bouquelet
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  3. M. Ali Pereyra
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  6. Raúl Chavez
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  9. Concepción Agundis
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Correspondence to Concepción Agundis.

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Solórzano, C., Bouquelet, S., Pereyra, M.A. et al. Isolation and characterization of the potential receptor for wheat germ agglutinin from human neutrophils. Glycoconj J 23, 591–598 (2006). https://doi.org/10.1007/s10719-006-8635-6

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  • DOI: https://doi.org/10.1007/s10719-006-8635-6

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