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'-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.
Ma
Jianjun
J
Division of Aquaculture and Biotechnology, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China.
Zhang
Dianchang
D
Jiang
Jingjing
J
Cui
Shuge
S
Pu
Hanlin
H
Jiang
Shigui
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D056668:Cathepsin L / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D004064:Digestive System / Q000276:immunology; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010802:Phylogeny; D049911:Pinctada / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D044149:Vibrio alginolyticus / Q000276:immunology
D056668:Cathepsin L