Molecular cloning, characterization and expression of cathepsin D from grass carp (Ctenopharyngodon idella)

Cathepsin D is a lysosomal aspartic proteinase which participates in various degradation functions within the cell. In this current study, we cloned and characterized the complete cDNA of grass carp cathepsin D through 5′- and 3′-RACE. The cathepsin D contained a 56 bp 5′ terminal untranslated region (5′-UTR), a 1197 bp open reading frame encoding 398 amino acids, and a 394 bp 3′-UTR. Grass carp cathepsin D shared high similarity with those from other species, and showed the highest amino acid identity of 91% to Danio rerio. Unlike many other organisms, the grass carp cathepsin D contains only one N-glycosylation site closest to the N-terminal. Real-time quantitative RT-PCR demonstrated that Cathepsin D expressed in all twelve tissues (bladder, brain, liver, heart, gill, muscle, fin, eye, intestines, spleen, gonad and head kidney). The relative expression levels of Cathepsin D in gonad and liver were 26.58 and 24.95 times as much as those in fin, respectively. The expression level of Cathepsin D in muscle approximately 16-fold higher, in intestines and spleen were 12-fold higher. The cathepsin D expression showed an upward trend during embryonic development. After challenged with Aeromonas hydrophil, the expression of grass carp cathepsin D gene showed significant changes in the four test tissues (liver, head kidney, spleen and intestines). The fact that the bacterial infection can obviously improve the cathepsin D expression in immune-related organs, may suggest that cathepsin D plays an important role in the innate immune response of grass carp.

► We cloned and characterized the complete cDNA of grass carp (Ctenopharyngodon idella) cathepsin D. ► The cathepsin D from grass carp has only one N-glycosylation site closest to the N-terminal. ► The cathepsin D expression showed an upward trend during embryonic development. ► Cathepsin D expressed in all twelve tissues, highest in gonad and liver, lowest in fin and brain. ► Challenge test may suggest that cathepsin D plays a key role in innate immune response of teleost.