TY - JOUR
T1 - Alpha2-macroglobulin from an Atlantic shrimp
T2 - Biochemical characterization, sub-cellular localization and gene expression upon fungal challenge
AU - Perazzolo, Luciane Maria
AU - Bachère, Evelyne
AU - Rosa, Rafael Diego
AU - Goncalves, Priscila
AU - Andreatta, Edemar Roberto
AU - Daffre, Sirlei
AU - Barracco, Margherita Anna
PY - 2011/12
Y1 - 2011/12
N2 - 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.
AB - 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.
KW - Alpha2-macroglobulin
KW - Farfantepenaeus paulensis
KW - Hemocyte
KW - Shrimp immunity
KW - Transcriptional response to fungal infection
UR - http://www.scopus.com/inward/record.url?scp=81255211056&partnerID=8YFLogxK
U2 - 10.1016/j.fsi.2011.08.011
DO - 10.1016/j.fsi.2011.08.011
M3 - Article
C2 - 21888978
AN - SCOPUS:81255211056
SN - 1050-4648
VL - 31
SP - 938
EP - 943
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
IS - 6
ER -