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Identification, mRNA expression profiling and activity characterization of cathepsin L from red drum (Sciaenops ocellatus)

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Abstract

Cathepsin L is a cysteine protease with a papain-like structure. It is known to be implicated in multiple processes of mammalian immune response to pathogen infection. In teleost fish, the functionality of cathepsin L is less understood. In this work, we characterized a cathepsin L homologue (designated as SoCatL) from red drum Sciaenops ocellatus, an important farmed fish species in China. SoCatL possesses a typical domain arrangement characteristic of cathepsin L, which comprises a proregion and a protease domain with four catalytically essential residues (Gln137, Cys143, His282 and Asn302) conserved in various organisms. SoCatL shares moderate sequence identities with mammalian cathepsin L and relatively high sequence identities with teleost cathepsin L. Phylogenetic analysis revealed that SoCatL is evolutionally close to fish cathepsin L, especially those belonging to the Perciformes order. The homology model of SoCatL was discovered to exhibit a structure resembling human cathepsin L. Transcriptional expression of SoCatL was found ubiquitous in tissues and enhanced after experimental infection with a bacterial pathogen. Recombinant SoCatL purified from Escherichia coli (designated as rSoCatL) displayed apparent proteolytic activity, which was optimal at 50 °C and pH 7.0. The activity of rSoCatL required the catalytic residue Cys143 and was severely reduced by cathepsin inhibitor. These results suggest that SoCatL is a teleost cathepsin L homologue which functions as a cysteine protease and is likely to participate in the host immune response against bacterial infection.

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Acknowledgments

This work was supported by the grants of the Chinese Ministry of Science and Technology (2012BAD17B01) and the Taishan Scholar Program of Shandong Province.

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  1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, China

    Bo-guang Sun & Yong-hua Hu

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  1. Bo-guang Sun
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Correspondence to Yong-hua Hu.

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Sun, Bg., Hu, Yh. Identification, mRNA expression profiling and activity characterization of cathepsin L from red drum (Sciaenops ocellatus). Fish Physiol Biochem 41, 1463–1473 (2015). https://doi.org/10.1007/s10695-015-0099-9

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  • DOI: https://doi.org/10.1007/s10695-015-0099-9

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