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Acta Parasitologica


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Volume 58, Issue 4

Issues

Trichomonas vaginalis acidic phospholipase A2: isolation and partial amino acid sequence

Brenda Escobedo-Guajardo
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
  • Departamento de Biología Celular y Genética, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León San Nicolás de los Garza, C.P. 66451, Nuevo León, México
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/ Francisco González-Salazar
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
  • Departamento de Ciencias Básicas, División de Ciencias de la Salud, Universidad de Monterrey, Av. Morones Prieto 4500 Pte. San Pedro Garza García, C.P. 66238, Nuevo León, México
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/ Rebeca Palacios-Corona
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
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/ Víctor Torres de la Cruz
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
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/ Mario Morales-Vallarta
  • Departamento de Biología Celular y Genética, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León San Nicolás de los Garza, C.P. 66451, Nuevo León, México
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/ Benito Mata-Cárdenas
  • Facultad de Ciencias Químicas, Universidad Autónoma de Nuevo León, Av. Manuel L Barragán S/N, San Nicolás de los Garza, Nuevo León, México
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/ Jesús Garza-González
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
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/ Gerardo Rivera-Silva
  • Departamento de Ciencias Básicas, División de Ciencias de la Salud, Universidad de Monterrey, Av. Morones Prieto 4500 Pte. San Pedro Garza García, C.P. 66238, Nuevo León, México
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/ Javier Vargas-Villarreal
  • División de Biología Celular y Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social. Administración de Correo No. 4, Apartado Postal 020-E, Colonia Independencia, Monterrey Nuevo León, México
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Published Online: 2013-12-13 | DOI: https://doi.org/10.2478/s11686-013-0166-2

Abstract

Sexually transmitted diseases are a major cause of acute disease worldwide, and trichomoniasis is the most common and curable disease, generating more than 170 million cases annually worldwide. Trichomonas vaginalis is the causal agent of trichomoniasis and has the ability to destroy in vitro cell monolayers of the vaginal mucosa, where the phospholipases A2 (PLA2) have been reported as potential virulence factors. These enzymes have been partially characterized from the subcellular fraction S30 of pathogenic T. vaginalis strains. The main objective of this study was to purify a phospholipase A2 from T. vaginalis, make a partial characterization, obtain a partial amino acid sequence, and determine its enzymatic participation as hemolytic factor causing lysis of erythrocytes. Trichomonas S30, RF30 and UFF30 sub-fractions from GT-15 strain have the capacity to hydrolyze [2-14C-PA]-PC at pH 6.0. Proteins from the UFF30 sub-fraction were separated by affinity chromatography into two eluted fractions with detectable PLA A2 activity. The EDTA-eluted fraction was analyzed by HPLC using on-line HPLC-tandem mass spectrometry and two protein peaks were observed at 8.2 and 13 kDa. Peptide sequences were identified from the proteins present in the eluted EDTA UFF30 fraction; bioinformatic analysis using Protein Link Global Server charged with T. vaginalis protein database suggests that eluted peptides correspond a putative ubiquitin protein in the 8.2 kDa fraction and a phospholipase preserved in the 13 kDa fraction. The EDTA-eluted fraction hydrolyzed [2-14C-PA]-PC lyses erythrocytes from Sprague-Dawley in a time and dose-dependent manner. The acidic hemolytic activity decreased by 84% with the addition of 100 μM of Rosenthal’s inhibitor.

Keywords: Trichomonas vaginalis; amino acid sequence; isolation; phospholipases; phospholipases A2; virulence factors

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About the article

Published Online: 2013-12-13

Published in Print: 2013-12-01


Citation Information: Acta Parasitologica, Volume 58, Issue 4, Pages 519–526, ISSN (Online) 1896-1851, ISSN (Print) 1230-2821, DOI: https://doi.org/10.2478/s11686-013-0166-2.

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© 2013 W. Stefański Institute of Parasitology, PAS. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

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