title: Alpha2-macroglobulin from an Atlantic shrimp: biochemical characterization, sub-cellular localization and gene expression upon fungal challenge.
abstract: In this study, we report on the isolation and characterization of an alpha2-macroglobulin (α2M) from the plasma of the pink shrimp Farfantepenaeus paulensis, its sub-cellular localization and transcriptional changes after infection by fungi. The molecular mass of the α2M was estimated at 389 kDa by gel filtration and 197 kDa by SDS-PAGE, under reducing conditions, suggesting that α2M from F. paulensis consists of two identical sub-units, covalently linked by disulphide bonds. The N-terminal amino acid sequence of the α2M from F. paulensis was very similar to those of other penaeid shrimps, crayfish and lobster (70-90% identity) and to a less extent with that of freshwater prawn (40% identity). A monoclonal antibody raised against the Marsupenaeus japonicus α2M made it possible to demonstrate that α2M of F. paulensis is stored in the vesicles of the shrimp granular hemocytes (through immunogold assay). Quantitative real-time PCR (qPCR) analysis showed that α2M mRNA transcripts significantly increased 24 h after an experimental infection with the shrimp pathogen Fusarium solani and it returned to the basal levels at 48 h post-injection. This is the first report on a α2M characterization in an Atlantic penaeid species and its expression profile upon a fungal infection.
authors: Laboratório de Imunologia Aplicada à Aquicultura, Departamento de Biologia Celular, Embriologia e Genética (BEG), Centro de Ciências Biológicas, Universidade Federal de Santa Catarina (UFSC), CEP 88040-900, Florianópolis, SC, Brazil. luciane@ccb.ufsc.br
authors: Luciane Maria
authors: LM
authors: Perazzolo
authors: Evelyne
authors: E
authors: Bachère
authors: Rafael Diego
authors: RD
authors: Rosa
authors: Priscila
authors: P
authors: Goncalves
authors: Edemar Roberto
authors: ER
authors: Andreatta
authors: Sirlei
authors: S
authors: Daffre
authors: Margherita Anna
authors: MA
authors: Barracco
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D005670:Fusarium / Q000276:immunology*; D005786:Gene Expression Regulation / Q000276:immunology*; D006434:Hemocytes / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000276:immunology* / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013045:Species Specificity; D000511:alpha-Macroglobulins / Q000235:genetics / Q000276:immunology*
chemical_list: D017931:DNA Primers; D000511:alpha-Macroglobulins