Characterization and functional analysis of fibrinogen-related protein (FreP) in the black tiger shrimp, Penaeus monodon. Issue 109 (February 2021)

• Record Type: Journal Article
• Authors: Oangkhana, Prawit; Amparyup, Piti; Tassanakajon, Anchalee; Preetham, Elumalai; Wongpanya, Ratree
• Is Part Of: Fish & shellfish immunology. Issue 109(2021)
• Page Start: 87
• Page End: 96
• Publication Date: 2021-02
• Subjects: FreD -- Fibrinogen-related domain -- Innate immunity -- Shrimp; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2020.12.012
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Ficolin is classified as an immune related protein containing collagen-like and fibrinogen-related domain (FreD). In invertebrates, the functions of fibrinogen-related proteins (FrePs) are of importance to innate immunity. In this study, a FreP in the black tiger shrimp Penaeus monodon was identified and characterized. The Pm FreP cDNA is 1, 007 bp long with a 921 bp-open reading frame that encodes for 306 amino acids. The deduced Pm FreP sequence consists of a signal peptide, an unknown region and the FreD. Phylogenetic analysis showed that Pm FreP was clustered with fibrinogen-like proteins in crustaceans which was separated from vertebrate ficolin-like proteins. The deduced fibrinogen-like domain contains four conserved cysteine residues (Cys96, Cys127, Cys249, and Cys262) that are responsible for the formation of disulfide bridges. Gene expression analysis shows that Pmfrep is mainly expressed in the intestine and the expression is significantly upregulated after Vibrio harveyi and white spot syndrome virus (WSSV) challenge. Recombinant Pm FreP (r Pm FreP) were successfully expressed and purified, and forms a trimeric structure as judged by native-PAGE. Bacterial binding assay showed that the r Pm FreD can bind and agglutinate Gram-negative and Gram-positive bacteria in the presence of calcium (Ca 2+ ) ions. Moreover, the r Pm FreP facilitates the clearance of V. harveyi in vivo . Overall, our results suggested that the Pm FreP may serve as pattern recognition receptors implicated in shrimp innate immunity. Highlights: Fibrinogen-related protein, Pm FreP, was cloned and characterized from Penaeus monodon. Pm FreP is mainly expressed in the intestine. Pm FreP is significantly upregulated after Vibrio harveyi and white spot syndrome virus (WSSV) challenge. r Pm FreP protein binds and aggregates Gram-positive and Gram-negative bacteria with clearance activity of V. harveyi in vivo.