title: Characterization of a lectin from the craysfish Cherax quadricarinatus hemolymph and its effect on hemocytes.
abstract: Lectins participate in the immune mechanisms of crustaceans. They have been considered as humoral receptors for pathogen-associated molecular patterns; however, some reports suggest that lectins could regulate crustacean cellular functions. In the present study, we purified and characterized a serum lectin (CqL) from the hemolymph of Cherax quadricarinatus by affinity chromatography and determined its participation in the regulation of hemocytes' oxidative burst. CqL is a 290-kDa lectin in native form, constituted by 108, 80, and 29-kDa subunits. It is mainly composed of glycine, alanine, and a minor proportion of methionine and histidine. It showed no carbohydrates in its structure. CqL is composed of several isoforms, as determined by 2D-electrophoresis, and shows no homology with any crustacean protein as determined by Lc/Ms mass spectrometry. CqL agglutinated mainly rat and rabbit erythrocytes and showed a broad specificity for monosaccharides such as galactose, glucose, and sialic acid, as well as for glycoproteins, such as porcine stomach and bovine submaxillary mucin and fetuin. It is a Mn(2+)-dependent lectin. CqL recognized 8% of crayfish granular hemocytes and increased 4.2-fold the production of hemocytes' superoxide anion in vitro assays when compared with non-treated hemocytes. This effect showed the same specificity for carbohydrates as hemagglutination; moreover, superoxide dismutase and diphenyleneiodonium chloride were effective inhibitors of CqL oxidative-activation. The CqL homoreceptor is a 120-kDa glycoprotein identified in the hemocytes lysate. Our results suggest that CqL participates actively in the regulation of the generation of superoxide anions in hemocytes using NADPH-dependent mechanisms.
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico; Posgrado de Ciencias del Mar y Limnologia, UNAM, 04510, Mexico.
authors.forename: J L
authors.initials: JL
authors.lastname: Sánchez-Salgado
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico.
authors.forename: M A
authors.initials: MA
authors.lastname: Pereyra
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico.
authors.forename: O
authors.initials: O
authors.lastname: Vivanco-Rojas
authors.affiliation: Centro de Investigaciones Biologicas, Universidad Autonoma del Estado de Morelos, Cuernavaca 62209, Mexico.
authors.forename: C
authors.initials: C
authors.lastname: Sierra-Castillo
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico; Instituto Tecnologico de Oaxaca, Oaxaca 68030, Mexico.
authors.forename: J J
authors.initials: JJ
authors.lastname: Alpuche-Osorno
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico; Centro de Investigaciones UNAM-UABJO, Oaxaca 68020, Mexico.
authors.forename: E
authors.initials: E
authors.lastname: Zenteno
authors.affiliation: Departamento de Bioquimica, Laboratorio de Inmunologia, Facultad de Medicina UNAM, 04510, Mexico. Electronic address: agundis@bq.unam.mx.
authors.forename: C
authors.initials: C
authors.lastname: Agundis
mesh_terms: D000372:Agglutination Tests; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003400:Astacoidea / Q000737:chemistry* / Q000276:immunology*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D006434:Hemocytes / Q000276:immunology* / Q000378:metabolism; D006458:Hemolymph / Q000378:metabolism; D007150:Immunohistochemistry; D037102:Lectins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D013058:Mass Spectrometry; D010587:Phagocytosis / Q000276:immunology; D017382:Reactive Oxygen Species / Q000378:metabolism; D016897:Respiratory Burst / Q000276:immunology; D018709:Statistics, Nonparametric
chemical_list: D000596:Amino Acids; D037102:Lectins; D017382:Reactive Oxygen Species
keywords: Cherax quadricarinatus; Crustacean lectin; Immune system; Lectin receptor; Oxidative burst