Inhibitory effect of a reproductive-related serpin on sperm trypsin-like activity implicates its role in sperm maturation of Penaeus monodon

In a number of marine animals, sperm serine proteases are important for fertilization. Penaeus monodon sperm require trypsin-like activity for a complete acrosome reaction, which exclusively occurs in sperm residing in the female thelycum. In this study, a complete cDNA sequence of reproductive tract-related Serine protease inhibitor (rrPmserpin) was identified. The longest open reading frame was composed of 1,366 nucleotides encoding 402 amino acids with a predicted pI of 6.86 and molecular mass of 44.88kDa. The signal peptide cleavage site was identified as the 17th amino acid residue in the amino-terminus, and two potential N-glycosylation sites were predicted as post-translation modifications. A conserved reactive loop and fold similarities, identified through three-dimensional modeling, suggested that this gene is a member of the serpin family. The expression of rrPmserpin mRNA was prominent in the reproductive organs, including the testis, vas deferens, terminal ampoule containing the spermatophore, and the female thelycum. Inhibitory activity of recombinant rrPmSERPIN-6His was revealed from the negative correlation between the abundance of rrPmserpin mRNA and sperm trypsin-like activities, along with its inhibitory effects on chymotrypsin, trypsin, and thelycal proteases. Therefore, our results suggest that rrPmserpin participates in the regulation of the activity of a sperm protease and the decapacitation process.