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Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum.
Mol Biochem Parasitol. 2008 Aug; 160(2):129-37.MB

Abstract

Gnathostoma spinigerum is a causative agent of human gnathostomiasis, a common parasitic disease involving skin and visceral organs, especially the central nervous system. In this study, we identified a cDNA encoding a cathepsin L-like cysteine protease (GsCL1) from the lambdaZAP cDNA library of G. spinigerum advanced third-stage larva (aL3) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1484bp encoded 398 amino acids which contained a typical signal peptide sequence (23 amino acids), a pro-domain (156 amino acids), and a mature domain (219 amino acids) with an approximate molecular weight of 24kDa. The deduced amino acid sequence of GsCL1 gene showed 53-64% identity to cathepsin L proteases of various organisms including a cathepsin L family member (cpl-1) of Caenorhabditis elegans. Recombinant proGsCL1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. The expressed enzyme displayed optimal protease activity toward Z-Phe-Arg-AMC substrate at pH 6.0 but not toward Z-Arg-Arg-AMC. The activity was sensitive to cysteine protease inhibitors E-64 and K11777. The preference for large hydrophilic and aromatic residues in the P2 position (I, L, F, W, U, V) was typical of cathepsin L proteases. Mouse anti-GST-proGsCL1 serum showed reactivity with 35-, 38- and 45-kDa proteins in the aL3 extracts. These proteins were shown to localize inside the intestinal cells of aL3.

Authors+Show Affiliations

Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18554733

Citation

Kongkerd, Natthawan, et al. "Identification and Characterization of a Cathepsin L-like Cysteine Protease From Gnathostoma Spinigerum." Molecular and Biochemical Parasitology, vol. 160, no. 2, 2008, pp. 129-37.
Kongkerd N, Uparanukraw P, Morakote N, et al. Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum. Mol Biochem Parasitol. 2008;160(2):129-37.
Kongkerd, N., Uparanukraw, P., Morakote, N., Sajid, M., & McKerrow, J. H. (2008). Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum. Molecular and Biochemical Parasitology, 160(2), 129-37. https://doi.org/10.1016/j.molbiopara.2008.05.001
Kongkerd N, et al. Identification and Characterization of a Cathepsin L-like Cysteine Protease From Gnathostoma Spinigerum. Mol Biochem Parasitol. 2008;160(2):129-37. PubMed PMID: 18554733.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum. AU - Kongkerd,Natthawan, AU - Uparanukraw,Pichart, AU - Morakote,Nimit, AU - Sajid,Mohammed, AU - McKerrow,James H, Y1 - 2008/05/08/ PY - 2008/02/02/received PY - 2008/05/01/revised PY - 2008/05/01/accepted PY - 2008/6/17/pubmed PY - 2008/9/13/medline PY - 2008/6/17/entrez SP - 129 EP - 37 JF - Molecular and biochemical parasitology JO - Mol Biochem Parasitol VL - 160 IS - 2 N2 - Gnathostoma spinigerum is a causative agent of human gnathostomiasis, a common parasitic disease involving skin and visceral organs, especially the central nervous system. In this study, we identified a cDNA encoding a cathepsin L-like cysteine protease (GsCL1) from the lambdaZAP cDNA library of G. spinigerum advanced third-stage larva (aL3) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1484bp encoded 398 amino acids which contained a typical signal peptide sequence (23 amino acids), a pro-domain (156 amino acids), and a mature domain (219 amino acids) with an approximate molecular weight of 24kDa. The deduced amino acid sequence of GsCL1 gene showed 53-64% identity to cathepsin L proteases of various organisms including a cathepsin L family member (cpl-1) of Caenorhabditis elegans. Recombinant proGsCL1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. The expressed enzyme displayed optimal protease activity toward Z-Phe-Arg-AMC substrate at pH 6.0 but not toward Z-Arg-Arg-AMC. The activity was sensitive to cysteine protease inhibitors E-64 and K11777. The preference for large hydrophilic and aromatic residues in the P2 position (I, L, F, W, U, V) was typical of cathepsin L proteases. Mouse anti-GST-proGsCL1 serum showed reactivity with 35-, 38- and 45-kDa proteins in the aL3 extracts. These proteins were shown to localize inside the intestinal cells of aL3. SN - 0166-6851 UR - https://www.unboundmedicine.com/medline/citation/18554733/Identification_and_characterization_of_a_cathepsin_L_like_cysteine_protease_from_Gnathostoma_spinigerum_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-6851(08)00115-1 DB - PRIME DP - Unbound Medicine ER -