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

for the Cysticercosis Working Group in Peru

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalVeterinary Parasitology
Volume267
DOIs
StatePublished - Mar 1 2019

Fingerprint

Taenia solium
cathepsin L
Cathepsin L
metacestodes
Cysticercosis
cysticercosis
Immunologic Tests
Cysteine Proteases
serodiagnosis
cysteine proteinases
Peptide Hydrolases
Swine
proteinases
zymogens
Enzyme Precursors
swine
Baculoviridae
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
peptides

Keywords

  • Baculovirus expression vector system
  • Cathepsin L
  • Cysticercosis
  • Immunodiagnosis
  • Taenia solium

ASJC Scopus subject areas

  • Parasitology
  • veterinary(all)

Cite this

Characterization of a novel cathepsin L-like protease from Taenia solium metacestodes for the immunodiagnosis of porcine cysticercosis. / for the Cysticercosis Working Group in Peru.

In: Veterinary Parasitology, Vol. 267, 01.03.2019, p. 9-16.

Research output: Contribution to journalArticle

@article{56b0e47ef5484befbbee290329b8a82d,
title = "Characterization of a novel cathepsin L-like protease from Taenia solium metacestodes for the immunodiagnosis of porcine cysticercosis",
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.",
keywords = "Baculovirus expression vector system, Cathepsin L, Cysticercosis, Immunodiagnosis, Taenia solium",
author = "{for the Cysticercosis Working Group in Peru} and Nancy Le{\'o}n-Janampa and Ruddy Liendo and Gilman, {Robert H} and Carlos Padilla and Garc{\'i}a, {Hector H.} and Armandoe Gonzalez and Patricia Sheen and Pajuelo, {M{\'o}nica J.} and Mirko Zimic",
year = "2019",
month = "3",
day = "1",
doi = "10.1016/j.vetpar.2019.01.004",
language = "English (US)",
volume = "267",
pages = "9--16",
journal = "Veterinary Parasitology",
issn = "0304-4017",
publisher = "Elsevier",

}

TY - JOUR

T1 - Characterization of a novel cathepsin L-like protease from Taenia solium metacestodes for the immunodiagnosis of porcine cysticercosis

AU - for the Cysticercosis Working Group in Peru

AU - León-Janampa, Nancy

AU - Liendo, Ruddy

AU - Gilman, Robert H

AU - Padilla, Carlos

AU - García, Hector H.

AU - Gonzalez, Armandoe

AU - Sheen, Patricia

AU - Pajuelo, Mónica J.

AU - Zimic, Mirko

PY - 2019/3/1

Y1 - 2019/3/1

N2 - 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.

AB - 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.

KW - Baculovirus expression vector system

KW - Cathepsin L

KW - Cysticercosis

KW - Immunodiagnosis

KW - Taenia solium

UR - http://www.scopus.com/inward/record.url?scp=85060855135&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060855135&partnerID=8YFLogxK

U2 - 10.1016/j.vetpar.2019.01.004

DO - 10.1016/j.vetpar.2019.01.004

M3 - Article

C2 - 30878092

AN - SCOPUS:85060855135

VL - 267

SP - 9

EP - 16

JO - Veterinary Parasitology

JF - Veterinary Parasitology

SN - 0304-4017

ER -