Cloning and characterization of a clip domain serine protease and its homolog (masquerade) from Eriocheir sinensis

• One cSP (EscSP) were identified from Eriocheir sinensis.
• One SPH, similar to Drosophila masquerade (EsMas), were also identified from E. sinensis.
• EscSP has only one clip domain, but EsMas has seven clip domains.
• EscSP in hemocytes showed significant increase after a LPS or PGN challenge.
• Down-regulation of EsMas challenged by LPS and EsMas could be induced by PGN challenge.

Serine proteinases (SPs) or SP homologs (SPHs) including clip domain SPs (cSPs) or SPHs (cSPHs) play critical roles in digestion, embryonic development, hemolymph coagulation, and melanization. In this study, one cSP (EscSP) and one SPH, similar to Drosophila masquerade (EsMas), were identified from hepatopancreas of the Chinese mittern crab Eriocheir sinensis. They both possess the clip domains at the N-terminal, EscSP has only one clip domain, but EsMas has seven clip domains. One SP or SP-like domain was at the C-terminal of EscSP and EsMas respectively. In contrast to EscSP, absence of a catalytic residue of Ser resulted in the loss of SP activity of EsMas. Tissue distribution analysis showed that EscSP mRNA was mainly expressed in hepatopancreas, nerve and eyestalk tissue; whereas the EsMas transcript was mainly distributed in eyestalk, muscle, nerve and hemocytes. EscSP in hemocytes showed significant increase after a lipopolysaccharide (LPS) or peptidoglycan (PGN) challenge. However, down-regulation of EsMas was observed in hemocytes challenged by LPS from 2 to 24 h, by contrast EsMas could be induced by PGN challenge at 2 and 24 h. All these findings indicated that EscSP and EsMas might be involved in the innate immune defenses in E. sinensis.