Molecular characterization of the cathepsin B of turbot (Scophthalmus maximus)

• Published in: Fish Physiology and Biochemistry
• Main Authors: Zhou, Ze-jun, Qiu, Reng, Zhang, Jian
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2015
• Subjects: Cathepsin b; Scophthalmus Maximus; Expression; Cysteine Protease; Scophthalmus maximus; Turbot
• Online Access: http://ir.qdio.ac.cn/handle/337002/25441; https://doi.org/10.1007/s10695-014-9998-4

Cathepsin B is an enzymatic protein belonging to the peptidase C1 family. It is involved in diverse physiological and pathological functions that include immune response. In this study, we identified and characterized a cathepsin B homolog (SmCatB) from turbot (Scophthalmus maximus). SmCatB is composed of 330 amino acid residues and possesses typical domain architecture of cathepsin B, which contains a propeptide region and a cysteine protease domain, and the latter processes four conserved residues (Q101, C107, H277, and N297) in the active site. SmCatB shares 80.6-87.6 % overall sequence identities with the cathepsin B of a number of teleost. SmCatB expression was detected in a wide range of tissues and upregulated by bacterial infection in a time-dependent manner. Recombinant SmCatB (rSmCatB-WT) purified from Escherichia coli exhibited apparent protease activity, which was optimal at 50 degrees C and pH 5.5. Compared to rSmCatB-WT, the mutant proteins rSmCatB-C107S, rSmCatB-H277A, and rSmCatB-N297A, which bear C107S, H277A, and N297A mutations, respectively, were significantly reduced in protease activity, with the highest reduction observed with rSmCatB-N297A. These results indicate that SmCatB is a bioactive protease that depends on the conserved structural features and that SmCatB is involved in pathogen-induced immune response.