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A murrel cysteine protease, cathepsin L: bioinformatics characterization, gene expression and proteolytic activity

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Abstract

Cathepsin L, a lysosomal endopeptidase, is a member of the peptidase C1 family (papain-like family) of cysteine proteinases that cleave peptide bonds of lysosomal proteins. In this study, we report a cathepsin L sequence identified from the constructed cDNA library of striped murrel Channa striatus (designated as CsCath L) using genome sequencing FLXTM technology. The full-length CsCath L contains three eukaryotic thiol protease domains at positions 134-145, 278-288 and 299-318. Phylogenetic analysis revealed that the CsCath L was clustered together with other cathepsin L from teleosts. The three-dimensional structure of CsCath L modelled by the I-Tasser program was compared with structures deposited in the Protein Data Bank to find out the structural similarity of CsCath L with experimentally identified structures. The results showed that the CsCath L exhibits maximum structural identity with pro-cathepsin L from human. The RNA fold structure of CsCath L was predicted along with its minimum free energy (−471.93 kcal/mol). The highest CsCath L gene expression was observed in liver, which was also significantly higher (P < 0.05) than that detected in other tissues taken for analysis. In order to investigate the mRNA transcription profile of CsCath L during infection, C. striatus were injected with fungus (Aphanomyces invadans) and bacteria (Aeromonas hydrophila) and its expression was up-regulated in liver at various time points. Similar to gene expression studies, the highest CsCath L enzyme activity was also observed in liver and its activity was up-regulated by fungal and bacterial infections.

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Abbreviations

CsCath L:

Channa striatus cathepsin L

EUS:

epizootic ulcerative syndrome

MFE:

minimum free energy

ORF:

open reading frame

PBS:

phosphate buffer saline

PDB:

Protein Data Bank

qRT-PCR:

quantitative real time polymerase chain reaction

UTR:

untranslated region

References

  • Ahn S., Sung J., Kim N., Lee A., Jeon S., Lee J., Kim J., Chung J. & Lee H. 2010. Molecular cloning, expression and characterization of cathepsin L from mud loach (Misgurnus mizolepis). Appl. Biochem. Biotechnol. 162: 1858–1871.

    Article  CAS  PubMed  Google Scholar 

  • Aranishi F., Ogata H., Hara K., Osatomi K. & Ishihara T. 1997. Purification and characterization of cathepsin L from hepatopancreas of carp Cyprinus carpio. Comp. Biochem. Physiol. B Biochem. Mol. 118: 531–537.

    Article  CAS  Google Scholar 

  • Arockiaraj J., Avin F.A., Vanaraja P., Easwvaran S., Singh A., Othman R.Y. & Bhassu S. 2012. Immune role of MrNFκBI-α, an IκB family member characterized in prawn M. rosenbergii. Fish Shellfish Immunol. 33: 619–625.

    Article  CAS  PubMed  Google Scholar 

  • Arockiaraj J., Gnanam A., Muthukrishnan D., Pasupuleti M., Milton J. & Singh A. 2013. An upstream initiator caspase 10 of snakehead murrel Channa striatus, containing DED, p20 and p10 subunits: molecular cloning, gene expression and proteolytic activity. Fish Shellfish Immunol. 34: 505–513.

    Article  CAS  PubMed  Google Scholar 

  • Arockiaraj J., Vanaraja P., Easwvaran S., Singh A., Alinejaid T., Othman R. & Bhassu S. 2011. Gene profiling and characterization of arginine kinase-1 (MrAK-1) from freshwater giant prawn (Macrobrachium rosenbergii). Fish Shellfish Immunol. 31: 81–89.

    Article  CAS  PubMed  Google Scholar 

  • Balaji K., Schaschke N., Machleidt W., Catalfamo M. & Henkart P.A. 2002. Surface cathepsin B protects cytotoxic lymphocytes from self-destruction after degranulation. J. Exp. Med. 196: 493–503.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Caster G. & Cole J.R. 1990. Diagnostic Procedure in Veterinary Bacteriology and Mycology, 5th Edn. Academic Press.

    Google Scholar 

  • Chen L., Zhang M. & Sun L. 2011. Identification and expressional analysis of two cathepsins from half-smooth tongue sole (Cynoglossus semilaevis). Fish Shell?sh Immunol. 31: 1270–1277.

    Article  CAS  Google Scholar 

  • Chevreux B., Pfisterer T., Drescher B., Driesel A., Müller W., Wetter T. & Suhai S. 2004. Using the miraEST assembler for reliable and automated mRNA transcript assembly and SNP detection in sequenced ESTs. Genome Res. 14: 1147–1159.

    Article  CAS  PubMed  Google Scholar 

  • Conus S. & Simon H. 2010. Cathepsins and their involvement in immune responses. Swiss Med. Wkly 140: w13042.

    PubMed  Google Scholar 

  • Coulombe R., Grochulski P., Sivaraman J., Menard R., Mort J. & Cygler M. 1996. Structure of human procathepsin L reveals the molecular basis of inhibition by the prosegment. EMBO J. 15: 5492–5503.

    CAS  PubMed  Google Scholar 

  • Dhanaraj M., Haniffa M., Ramakrishnan C. & Singh S.V. 2008. Microbial flora from the Epizootic Ulcerative Syndrome (EUS) infected murrel Channa striatus (Bloch, 1797) in Tirunelveli region. Turk. J. Vet. Anim. Sci. 32: 221–224.

    Google Scholar 

  • Dong W., Xiang L. & Shao J. 2007. Cloning and characterisation of two natural killer enhancing factor genes (NKEF-A and NKEF-B) in pufferfish, Tetraodon nigroviridis. Fish Shellfish Immunol. 22: 1–15.

    Article  CAS  PubMed  Google Scholar 

  • Felbor U., Kessler B., Mothes W., Goebel H., Ploegh H., Bronson R. & Olsen B. 2002. Neuronal loss and brain atrophy in mice lacking cathepsins B and L. Proc. Natl. Acad. Sci. USA 99: 7883–7888.

    Article  CAS  PubMed  Google Scholar 

  • Hall T. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acid Symp. Ser. 41: 95–98.

    CAS  Google Scholar 

  • Heu M., Kim H., Cho D., Godber J. & Pyeun J. 1997. Purification and characterization of cathepsin L-like enzyme from the muscle of anchovy, Engraulis japonica. Comp. Biochem. Physiol. B Biochem. Mol. 118: 523–529.

    Article  CAS  Google Scholar 

  • Karrer K., Peiffer S. & Ditomas M. 1993. Two distinct gene subfamilies within the family of cysteine protease genes. Proc. Natl. Acad. Sci. USA 90: 3063–3067.

    Article  CAS  PubMed  Google Scholar 

  • Kim J., Jeong J., Park H., Kim E., Kim H., Chae Y., Kim D. & Park C. 2011. Molecular identification and expression analysis of cathepsins O and S from rock bream, Oplegnathus fasciatus. Fish Shellfish Immunol. 31: 578–587.

    Article  CAS  PubMed  Google Scholar 

  • Lee J., Chen H. & Jiang S. 1993. Purification and characterization of proteinases identified as cathepsins L and L-like (58 kDa) proteinase from mackerel (Scomber australasicus). Biosci. Biotechnol. Biochem. 57: 1470–1476.

    Article  CAS  Google Scholar 

  • Livak K. & Schmittgen T. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 −ΔΔCT method. Methods 25: 402–408.

    Article  CAS  PubMed  Google Scholar 

  • Li W., Jin X., He L., Jiang H., Gong Y., Xie Y. & Wang Q. 2010. Molecular cloning, characterization, expression and activity analysis of cathepsin L in Chinese mitten crab, Eriocheir sinensis. Fish Shellfish Immunol. 29: 1010–1018.

    Article  CAS  PubMed  Google Scholar 

  • Li W.W., He L., Jin X.K., Jiang H., Chen L.L., Wang Y. & Wang Q. 2011. Molecular cloning, characterization and expression analysis of cathepsin A gene in Chinese mitten crab, Eriocheir sinensis. Peptides 32: 518–525.

    Article  CAS  PubMed  Google Scholar 

  • Ma J., Zhang D., Jiang J., Cui S., Pu H. & Jiang S. 2010. Molecular characterization and expression analysis of cathepsin L1 cysteine protease from pearl oyster Pinctada fucata. Fish Shellfish Immunol. 29: 501–507.

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto I., Watanabe H., Abe K., Arai S. & Emori Y. 1995. A putative digestive cysteine proteinase from Drosophila melanogaster is predominantly expressed in the embryonic and larval midgut. Eur. J. Biochem. 227: 582–587.

    Article  CAS  PubMed  Google Scholar 

  • Menard R. & Storer A. 1992. Oxyanion hole interactions in serine and cysteine proteases. Biol. Chem. 373: 393–400.

    CAS  Google Scholar 

  • Press C.M. & Evensen O. 1999. The morphology of the immune system in teleost fishes. Fish Shellfish Immunol. 9: 309–318.

    Article  Google Scholar 

  • Rawlings N., Morton F. & Barrett A. 2006. MEROPS: the peptidase database. Nucleic Acids Res. 34: D270–D272.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Reinheckel T., Deussing J., Roth W. & Peters C. 2001. Towards specific functions of lysosomal cysteine peptidases: phenotypes of mice deficient for cathepsin B or cathepsin L. Biol. Chem. 382: 735–741.

    CAS  PubMed  Google Scholar 

  • Roy A., Kucukural A. & Zhang Y. 2010. I-TASSER: a unified platform for automated protein structure and function prediction. Nat. Protocols 5: 725–738.

    Article  CAS  Google Scholar 

  • Stephens A., Rojo L., Araujo-Bernal S., Garcia-Carreno F. & Muhlia-Almazan A. 2012. Cathepsin B from the white shrimp Litopenaeus vannamei: cDNA sequence analysis, tissuesspecific expression and biological activity. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 161: 32–40.

    Article  CAS  PubMed  Google Scholar 

  • Tamura K., Peterson D., Peterson N., Stecher G., Nei M. & Kumar S. 2011. MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 2731–2739.

    Article  CAS  PubMed  Google Scholar 

  • Tingaud-Sequeira A. & Cerda J. 2007. Phylogenetic relationships and gene expression pattern of three different cathepsin L (Ctsl) isoforms in zebrafish: Ctsla is the putative yolk processing enzyme. Gene 386: 98–106.

    Article  CAS  PubMed  Google Scholar 

  • Trent M., Stead C., Tran A. & Hankins J. 2006. Diversity of endotoxin and its impact on pathogenesis. J. Endotoxin Res. 12: 205–223.

    Article  CAS  PubMed  Google Scholar 

  • Turk V., Turk B. & Turk D. 2001. Lysosomal cysteine proteases: facts and opportunities. EMBO J. 20: 4629–4633.

    Article  CAS  PubMed  Google Scholar 

  • Uinuk-Ool T., Takezaki N., Kuroda N., Figueroa F., Sato A., Samonte I., Mayer W. & Klein J. 2003. Phylogeny of antigenprocessing enzymes: cathepsins of a cephalochordate, an agnathan and a bony fish. Scand. J. Immunol. 58: 436–448.

    Article  CAS  PubMed  Google Scholar 

  • Vernet T., Berti P.J., De Montigny C., Musil R., Tessier D.C., Menard R., Magny M.C., Storer A.C. & Thomas D.Y. 1995. Processing of the papain precursor. The ionization of a conserved amino acid motif within the pro region participates in the regulation of intramolecular processing. J. Biol. Chem. 270: 10838–10846.

    Article  CAS  PubMed  Google Scholar 

  • Visessanguan W., Benjakul S. & An H. 2003. Purification and characterization of cathepsin L in arrowtooth flounder (Atheresthes stomias) muscle. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 134: 477–487.

    Article  PubMed  Google Scholar 

  • Yamashita M. & Konagaya S. 1990. Purification and characterization of cathepsin L from the white muscle of chum salmon, Oncorhynchus keta. Comp. Biochem. Physiol. B 96: 247–252.

    CAS  PubMed  Google Scholar 

  • Yeh H. & Klesius P. 2009. Channel catfish, Ictalurus punctatus, cysteine proteinases: cloning, characterisation and expression of cathepsin H and L. Fish Shellfish Immunol. 26: 332–338.

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y. 2008. I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 9: 40.

    Article  PubMed Central  PubMed  Google Scholar 

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Authors and Affiliations

  1. Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, 603 203, Chennai, Tamil Nadu, India

    Venkatesh Kumaresan, Prasanth Bhatt, Rajesh Palanisamy & Jesu Arockiaraj

  2. Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX, 78712, USA

    Annie J. Gnanam

  3. Lab PCN 206, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226 031, Uttar Pradesh, India

    Mukesh Pasupuleti

Authors
  1. Venkatesh Kumaresan
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  2. Prasanth Bhatt
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  3. Rajesh Palanisamy
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  4. Annie J. Gnanam
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  5. Mukesh Pasupuleti
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  6. Jesu Arockiaraj
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Correspondence to Jesu Arockiaraj.

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Kumaresan, V., Bhatt, P., Palanisamy, R. et al. A murrel cysteine protease, cathepsin L: bioinformatics characterization, gene expression and proteolytic activity. Biologia 69, 395–406 (2014). https://doi.org/10.2478/s11756-013-0326-8

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  • DOI: https://doi.org/10.2478/s11756-013-0326-8

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