Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum

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 λZAP cDNA library of G. spini...

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Main Authors: Kongkerd N., Uparanukraw P., Morakote N., Sajid M., McKerrow J.H.
Format: Article
Language:English
Published: 2014
Online Access:http://www.scopus.com/inward/record.url?eid=2-s2.0-50049131642&partnerID=40&md5=9884a2bdbf1e0d01261d1d7cfb97a09a
http://www.ncbi.nlm.nih.gov/pubmed/18554733
http://cmuir.cmu.ac.th/handle/6653943832/2398
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spelling th-cmuir.6653943832-23982014-08-30T02:00:48Z Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum Kongkerd N. Uparanukraw P. Morakote N. Sajid M. McKerrow J.H. 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 λZAP cDNA library of G. spinigerum advanced third-stage larva (aL3) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1484 bp 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 24 kDa. 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. © 2008 Elsevier B.V. 2014-08-30T02:00:48Z 2014-08-30T02:00:48Z 2008 Article 01666851 10.1016/j.molbiopara.2008.05.001 18554733 MBIPD http://www.scopus.com/inward/record.url?eid=2-s2.0-50049131642&partnerID=40&md5=9884a2bdbf1e0d01261d1d7cfb97a09a http://www.ncbi.nlm.nih.gov/pubmed/18554733 http://cmuir.cmu.ac.th/handle/6653943832/2398 English
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
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description 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 λZAP cDNA library of G. spinigerum advanced third-stage larva (aL3) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1484 bp 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 24 kDa. 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. © 2008 Elsevier B.V.
format Article
author Kongkerd N.
Uparanukraw P.
Morakote N.
Sajid M.
McKerrow J.H.
spellingShingle Kongkerd N.
Uparanukraw P.
Morakote N.
Sajid M.
McKerrow J.H.
Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
author_facet Kongkerd N.
Uparanukraw P.
Morakote N.
Sajid M.
McKerrow J.H.
author_sort Kongkerd N.
title Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
title_short Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
title_full Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
title_fullStr Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
title_full_unstemmed Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum
title_sort identification and characterization of a cathepsin l-like cysteine protease from gnathostoma spinigerum
publishDate 2014
url http://www.scopus.com/inward/record.url?eid=2-s2.0-50049131642&partnerID=40&md5=9884a2bdbf1e0d01261d1d7cfb97a09a
http://www.ncbi.nlm.nih.gov/pubmed/18554733
http://cmuir.cmu.ac.th/handle/6653943832/2398
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