title: Molecular characterization and expression analysis of Cathepsin B and L cysteine proteases from rock bream (Oplegnathus fasciatus).
abstract: Cathepsins are lysosomal cysteine proteases of the papain family that play an important role in intracellular protein degradation and turn over within the lysosomal system. In the present study, full-length sequences of cathepsin B (RbCathepsin B) and L (RbCathepsin L) were identified after transcriptome sequencing of rock bream Oplegnathus fasciatus mixed tissue cDNA. Cathepsin B was composed of 330 amino acid residues with 36 kDa predicted molecular mass. RbCathepsin L contained 336 amino acid residues encoding for a 38 kDa predicted molecular mass protein. The sequencing analysis results showed that both cathepsin B and L contain the characteristic papain family cysteine protease signature and active sites for the eukaryotic thiol proteases of cysteine, asparagine and histidine. In addition, RbCathepsin L contained EF hand Ca(2+) binding and cathepsin propeptide inhibitor domains. The rock bream cathepsin B and L showed the highest amino acid identity of 90 and 95% to Lutjanus argentimaculatus cathepsin B and Lates calcarifer cathepsin L, respectively. By phylogenetic analysis, cathepsin B and L exhibited a high degree of evolutionary relationship to respective cathepsin family members of the papain superfamily. Quantitative real-time RT-PCR analysis results confirmed that the expression of cathepsin B and L genes was constitutive in all examined tissues isolated from un-induced rock bream. Moreover, activation of RbCathepsin B and L mRNA was observed in both lipopolysaccharide (LPS) and Edwardsiella tarda challenged liver and blood cells, indicating a role of immune response in rock bream.
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
authors.forename: Ilson
authors.initials: I
authors.lastname: Whang
authors.forename: Mahanama
authors.initials: M
authors.lastname: De Zoysa
authors.forename: Chamilani
authors.initials: C
authors.lastname: Nikapitiya
authors.forename: Youngdeuk
authors.initials: Y
authors.lastname: Lee
authors.forename: Yucheol
authors.initials: Y
authors.lastname: Kim
authors.forename: Sukkyoung
authors.initials: S
authors.lastname: Lee
authors.forename: Chulhong
authors.initials: C
authors.lastname: Oh
authors.forename: Sung-Ju
authors.initials: SJ
authors.lastname: Jung
authors.forename: Myung-Joo
authors.initials: MJ
authors.lastname: Oh
authors.forename: Cheol Young
authors.initials: CY
authors.lastname: Choi
authors.forename: Sang-Yeob
authors.initials: SY
authors.lastname: Yeo
authors.forename: Bong-Seok
authors.initials: BS
authors.lastname: Kim
authors.forename: Se-Jae
authors.initials: SJ
authors.lastname: Kim
authors.forename: Jehee
authors.initials: J
authors.lastname: Lee
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002401:Cathepsin B / Q000235:genetics* / Q000378:metabolism*; D056668:Cathepsin L / Q000235:genetics* / Q000378:metabolism*; D020609:Edwardsiella tarda / Q000502:physiology; D004756:Enterobacteriaceae Infections / Q000201:enzymology / Q000276:immunology / Q000662:veterinary; D005393:Fish Diseases / Q000201:enzymology / Q000276:immunology; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D010473:Perciformes / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D016415:Sequence Alignment
chemical_list: D000276:Adjuvants, Immunologic; D008070:Lipopolysaccharides; D002401:Cathepsin B; D056668:Cathepsin L