Molecular characterization and expression analysis of cathepsin L1 cysteine protease from pearl oyster Pinctada fucata
Cathepsin L is one of the crucial enzyme superfamilies and involved in the immune responses. In this study, a cDNA encoding cathepsin L cysteine protease was identified and characterized from pearl oyster Pinctada fucata (designated as poCL1). The poCL1 cDNA was 1160 bp long and consisted of a 5′-un...
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Veröffentlicht in: | Fish & shellfish immunology 2010-09, Vol.29 (3), p.501-507 |
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Zusammenfassung: | Cathepsin L is one of the crucial enzyme superfamilies and involved in the immune responses. In this study, a cDNA encoding cathepsin L cysteine protease was identified and characterized from pearl oyster
Pinctada fucata (designated as poCL1). The poCL1 cDNA was 1160
bp long and consisted of a 5′-untranslated region (UTR) of 15
bp, a 3′-UTR of 149
bp with a polyadenylation signal (AATAAA) at 11 nucleotides upstream of the poly(A) tail, and an open reading frame (ORF) of 996
bp encoding a polypeptide of 331 amino acids, which contained a typical signal peptide sequence (Met
1–Ala
16), a prodomain (Thr
17–Asp
113), and a mature domain (Leu
114–Val
331). The preproprotein contained the oxyanion hole (Gln), the active triad formed by Cys, His and Asn, and the conserved ERFNIN, GNFD motifs, which is characteristic for cathepsin L proteases. Homology analysis revealed that the poCL1 shared 62.5–72.5% similarity and 42.9–56.0% identity to other known cathepsin L sequences. The phylogenetic tree showed that the poCL1 clustered with the invertebrate cathepsin L cysteine proteases and was closely related to
Stichopus japonicus CL,
Strongylocentrotus salar CL1 and
Radix peregra CL. The mRNA expression of the poCL1 in blank group and bacterial challenge group could be detected in all studied tissues with the higher level in digestive gland. The expression level of poCL1 mRNA was significantly up-regulated at 4
h and 8
h, and then significantly down-regulated at 12
h and 24
h in digestive gland after
Vibrio alginolyticus stimulation. These results provided important information for further exploring the roles of pearl oyster cathepsin L in the immune responses. |
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ISSN: | 1050-4648 1095-9947 |
DOI: | 10.1016/j.fsi.2010.05.006 |