Three clip domain serine proteases (cSPs) and one clip domain serine protease homologue (cSPH) identified from haemocytes and eyestalk cDNA libraries of swimming crab Portunus trituberculatus

Four genes including three clip domain serine proteases (PtcSP1, PtcSP2 and PtcSP3) and one clip domain serine protease homologue (PtcSPH) of the swimming crab Portunus trituberculatus (Decapoda: Brachyura: Portunidae) were characterized based on analysis of expressed sequence tags from haemocytes and eyestalk cDNA libraries. The relative four peptidases, which share high structural similarity to the clip-SPs of other arthropod species, appeared to possess a clip domain at the N-terminus and an enzymatically active serine protease domain at the C-terminus except PtcSPH for its second catalytic residue Asp. (D) replaced by Ala (A). Alignment among the four full-sequences showed that PtcSP2 and PtcSP3 had the highest identical score (58%) while the similarity of other sequences was lower than 24%. The mRNA transcripts of PtcSPs and PtcSPH could be detected widely in all the examined tissues with remarkable different expression levels. The temporal expressions of PtcSPs and PtcSPH demonstrated different time-dependent expression pattern post Vibrio alginolyticus, Micrococcus luteus, and Pichia pastoris challenge. Especially, the expression of PtcSPH transcripts showed greater change against V. alginolyticus compared with the other two microorganisms. These findings suggest that PtcSPs and PtcSPH play different roles in the antibacterial defence mechanism of P. trituberculatus crab.

► Four genes of Portunus trituberculatus (PtcSPH, PtcSP1-3) were characterized. ► The mRNA showed remarkable different expression levels in the examined tissues. ► The expression of PtcSPH showed greater change against Vibrio alginolyticus. ► PtcSPs and PtcSPH may play different roles in the antibacterial defence mechanism.