Related Experiment Video
Updated: May 29, 2026
Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium
Published on: April 9, 2019
Alpha2-macroglobulin from an Atlantic shrimp: biochemical characterization, sub-cellular localization and gene
Luciane Maria Perazzolo1, Evelyne Bachère, Rafael Diego Rosa
1Laboratório 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
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.
Insights
Researchers isolated and characterized alpha2-macroglobulin (α2M) from pink shrimp. Shrimp α2M levels increased after fungal infection, indicating its role in immune response.
Area of Science:
- Marine Biology
- Immunology
- Biochemistry
Background:
- Alpha2-macroglobulin (α2M) is a key immune protein in many species.
- Understanding shrimp immune responses is crucial for aquaculture.
- The Atlantic pink shrimp (Farfantepenaeus paulensis) is an important commercial species.
Purpose of the Study:
- To isolate and characterize α2M from Farfantepenaeus paulensis.
- To determine the sub-cellular localization of α2M in shrimp.
- To investigate the transcriptional changes of α2M following fungal infection.
Main Methods:
- Gel filtration and SDS-PAGE for molecular mass determination.
- N-terminal amino acid sequencing for homology analysis.
- Immunogold assay using a monoclonal antibody for sub-cellular localization.
- Quantitative real-time PCR (qPCR) for gene expression analysis.
Main Results:
- The molecular mass of F. paulensis α2M was estimated at 389 kDa (gel filtration) and 197 kDa (SDS-PAGE), suggesting a dimeric structure.
- N-terminal sequence showed high similarity to other penaeid shrimps.
- α2M was localized in the vesicles of granular hemocytes.
- α2M mRNA levels significantly increased 24 hours post-infection with Fusarium solani, returning to baseline by 48 hours.
Conclusions:
- This study provides the first characterization of α2M in an Atlantic penaeid species.
- The findings reveal α2M's localization in shrimp hemocytes and its upregulation during fungal infection.
- α2M plays a significant role in the innate immune response of Farfantepenaeus paulensis to fungal pathogens.
