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Characterization of a novel cathepsin L-like protease from Taenia solium metacestodes for the immunodiagnosis of porcine cysticercosis

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dc.contributor.author Leon-Janampa, Nancy
dc.contributor.author Liendo, Ruddy
dc.contributor.author Gilman, Robert Hugh
dc.contributor.author Padilla, Carlos
dc.contributor.author García Lescano, Héctor Hugo
dc.contributor.author Gonzales, Armando
dc.contributor.author Sheen Cortavarria, Patricia
dc.contributor.author Pajuelo Travezaño, Monica Jhenny
dc.contributor.author Zimic-Peralta, Mirko Juan
dc.contributor.author Cysticercosis Working Group in Peru
dc.date.accessioned 2019-07-04T16:59:24Z
dc.date.available 2019-07-04T16:59:24Z
dc.date.issued 2019
dc.identifier.uri https://hdl.handle.net/20.500.12866/6744
dc.description.abstract Porcine cysticercosis is an endemic parasitic disease caused by infection with Taenia solium that is found predominantly in developing countries. In order to aid in the development of simple diagnostic approaches, identification and characterization of potential new antigens for immunodiagnostic purposes is desired. The cysteine protease family has previously been found to have important immunodiagnostic properties. These proteases are expressed as zymogens which contain a signal peptide, pro-peptide, and an active domain. Subsequent catalytic cleavage of the pro-peptide converts these zymogens into enzymes. With the use of bioinformatic tools we identified an active domain of a novel cathepsin L-like cysteine protease (TsolCL) in the T. solium genome. The TsolCL gene includes 705 nucleotides (nt) within a single intron and a 633 nt exonic sequence encoding an active protein of 211 amino acids. Sequence alignment and phylogenetic analysis suggest that the TsolCL gene is closely related to genes found in Echinoccocus granulosus and E. multiloculars. In addition, TsolCL was found to have a 61.9%–99.0% similarity to other cathepsin L proteins found in other helminths and mammals. We cloned, expressed, purified, and characterized the recombinant active TsolCL (27 kDa) using the baculovirus-insect cell expression system. TsolCL showed cysteine protease enzymatic activity with the capacity to hydrolyze the Z-Phe-Arg-AMC substrate as well as bovine serum albumin. However, TsolCL was not able to hydrolyze human immunoglobulin. In addition, TsolCL has cathepsin L conserved amino acid residues in the catalytic site (Gln8, Cys14, His159, Asn179 and Trp181) and the motif GCNGG. Using ELISA, TsolCL was able to distinguish circulating IgG antibodies between healthy animals and naturally infected pigs with cysticercosis, showing a moderate sensitivity of 83.33% (40/48; 95% CI: [69.8%–92.5 %]), and a specificity of 83.78% (31/37; 95% CI: [67.9%–93.8%]). In conclusion, a novel cathepsin L-like cysteine protease from a T. solium metacestode was expressed successfully in Baculovirus system and was evaluated as a candidate antigen to diagnose porcine cysticercosis using the ELISA immunoassay. en_US
dc.language.iso eng
dc.publisher Elsevier
dc.relation.ispartofseries Veterinary Parasitology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject amino acid en_US
dc.subject amino acid sequence en_US
dc.subject animal en_US
dc.subject animal cell en_US
dc.subject animal experiment en_US
dc.subject animal model en_US
dc.subject Animals en_US
dc.subject Antibodies, Helminth en_US
dc.subject antigenicity en_US
dc.subject Antigens, Helminth en_US
dc.subject area under the curve en_US
dc.subject arginine en_US
dc.subject Article en_US
dc.subject aspartic acid en_US
dc.subject bacterial genome en_US
dc.subject Baculoviridae en_US
dc.subject Baculovirus expression vector system en_US
dc.subject bioinformatics en_US
dc.subject blood en_US
dc.subject bovine serum albumin en_US
dc.subject cathepsin L en_US
dc.subject Cathepsin L en_US
dc.subject cloning en_US
dc.subject controlled study en_US
dc.subject cysteine en_US
dc.subject cysticercosis en_US
dc.subject Cysticercosis en_US
dc.subject Echinococcus granulosus en_US
dc.subject Echinococcus multilocularis en_US
dc.subject enzyme activity en_US
dc.subject enzyme linked immunosorbent assay en_US
dc.subject enzyme substrate en_US
dc.subject Enzyme-Linked Immunosorbent Assay en_US
dc.subject enzymology en_US
dc.subject exon en_US
dc.subject gene expression en_US
dc.subject gene function en_US
dc.subject genetic similarity en_US
dc.subject genetic transfection en_US
dc.subject genetics en_US
dc.subject glycine en_US
dc.subject helminth en_US
dc.subject helminth antibody en_US
dc.subject histidine en_US
dc.subject hydrolysis en_US
dc.subject Immunodiagnosis en_US
dc.subject immunoglobulin en_US
dc.subject immunoglobulin G en_US
dc.subject Immunoglobulin G en_US
dc.subject immunoglobulin G antibody en_US
dc.subject Immunologic Tests en_US
dc.subject immunological procedures en_US
dc.subject insect cell en_US
dc.subject intron en_US
dc.subject isolation and purification en_US
dc.subject mammal en_US
dc.subject nonhuman en_US
dc.subject nucleotide en_US
dc.subject parasite antigen en_US
dc.subject parasitology en_US
dc.subject phenylalanine en_US
dc.subject phylogeny en_US
dc.subject Phylogeny en_US
dc.subject pig en_US
dc.subject predictive value en_US
dc.subject protein domain en_US
dc.subject protein motif en_US
dc.subject purification en_US
dc.subject receiver operating characteristic en_US
dc.subject recombinant protein en_US
dc.subject Recombinant Proteins en_US
dc.subject sensitivity and specificity en_US
dc.subject sequence alignment en_US
dc.subject sequence analysis en_US
dc.subject serodiagnosis en_US
dc.subject Sf9 cell line en_US
dc.subject Sf9 Cells en_US
dc.subject Swine en_US
dc.subject swine disease en_US
dc.subject Swine Diseases en_US
dc.subject Taenia solium en_US
dc.subject trypsin en_US
dc.subject veterinary medicine en_US
dc.subject virus recombinant en_US
dc.title Characterization of a novel cathepsin L-like protease from Taenia solium metacestodes for the immunodiagnosis of porcine cysticercosis en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.vetpar.2019.01.004
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.07
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#4.03.00
dc.relation.issn 1873-2550


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