Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence

Brenda L. Escobedo-Guajardo, Francisco González-Salazar, Rebeca Palacios-Corona, Víctor M. Torres de la Cruz, Mario Morales-Vallarta, Benito D. Mata-Cárdenas, Jesús N. Garza-González, Gerardo Rivera-Silva, Javier Vargas-Villarreal

Research output: Contribution to journalArticle

1 Citation (Scopus)

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. © 2013 Versita Warsaw and Springer-Verlag Wien.
Original languageEnglish
Pages (from-to)519-526
Number of pages8
JournalActa Parasitologica
DOIs
Publication statusPublished - 23 Dec 2013
Externally publishedYes

Fingerprint

Trichomonas vaginalis
Phospholipases A2
Amino Acid Sequence
Edetic Acid
Proteins
Erythrocytes
High Pressure Liquid Chromatography
Trichomonas
Protein Databases
Peptides
Subcellular Fractions
Phospholipases
Acute Disease
Virulence Factors
Sexually Transmitted Diseases
Ubiquitin
Tandem Mass Spectrometry
Computational Biology
Affinity Chromatography
varespladib methyl

All Science Journal Classification (ASJC) codes

  • Parasitology

Cite this

Escobedo-Guajardo, B. L., González-Salazar, F., Palacios-Corona, R., Torres de la Cruz, V. M., Morales-Vallarta, M., Mata-Cárdenas, B. D., ... Vargas-Villarreal, J. (2013). Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence. Acta Parasitologica, 519-526. https://doi.org/10.2478/s11686-013-0166-2
Escobedo-Guajardo, Brenda L. ; González-Salazar, Francisco ; Palacios-Corona, Rebeca ; Torres de la Cruz, Víctor M. ; Morales-Vallarta, Mario ; Mata-Cárdenas, Benito D. ; Garza-González, Jesús N. ; Rivera-Silva, Gerardo ; Vargas-Villarreal, Javier. / Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence. In: Acta Parasitologica. 2013 ; pp. 519-526.
@article{a0980c012f674b93ad60a21964a84318,
title = "Trichomonas vaginalis acidic phospholipase A2: Isolation and partial amino acid sequence",
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. {\circledC} 2013 Versita Warsaw and Springer-Verlag Wien.",
author = "Escobedo-Guajardo, {Brenda L.} and Francisco Gonz{\'a}lez-Salazar and Rebeca Palacios-Corona and {Torres de la Cruz}, {V{\'i}ctor M.} and Mario Morales-Vallarta and Mata-C{\'a}rdenas, {Benito D.} and Garza-Gonz{\'a}lez, {Jes{\'u}s N.} and Gerardo Rivera-Silva and Javier Vargas-Villarreal",
year = "2013",
month = "12",
day = "23",
doi = "10.2478/s11686-013-0166-2",
language = "English",
pages = "519--526",
journal = "Acta Parasitologica",
issn = "1230-2821",
publisher = "Walter de Gruyter GmbH",

}

Escobedo-Guajardo, BL, González-Salazar, F, Palacios-Corona, R, Torres de la Cruz, VM, Morales-Vallarta, M, Mata-Cárdenas, BD, Garza-González, JN, Rivera-Silva, G & Vargas-Villarreal, J 2013, 'Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence', Acta Parasitologica, pp. 519-526. https://doi.org/10.2478/s11686-013-0166-2

Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence. / Escobedo-Guajardo, Brenda L.; González-Salazar, Francisco; Palacios-Corona, Rebeca; Torres de la Cruz, Víctor M.; Morales-Vallarta, Mario; Mata-Cárdenas, Benito D.; Garza-González, Jesús N.; Rivera-Silva, Gerardo; Vargas-Villarreal, Javier.

In: Acta Parasitologica, 23.12.2013, p. 519-526.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Trichomonas vaginalis acidic phospholipase A2: Isolation and partial amino acid sequence

AU - Escobedo-Guajardo, Brenda L.

AU - González-Salazar, Francisco

AU - Palacios-Corona, Rebeca

AU - Torres de la Cruz, Víctor M.

AU - Morales-Vallarta, Mario

AU - Mata-Cárdenas, Benito D.

AU - Garza-González, Jesús N.

AU - Rivera-Silva, Gerardo

AU - Vargas-Villarreal, Javier

PY - 2013/12/23

Y1 - 2013/12/23

N2 - 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. © 2013 Versita Warsaw and Springer-Verlag Wien.

AB - 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. © 2013 Versita Warsaw and Springer-Verlag Wien.

U2 - 10.2478/s11686-013-0166-2

DO - 10.2478/s11686-013-0166-2

M3 - Article

SP - 519

EP - 526

JO - Acta Parasitologica

JF - Acta Parasitologica

SN - 1230-2821

ER -

Escobedo-Guajardo BL, González-Salazar F, Palacios-Corona R, Torres de la Cruz VM, Morales-Vallarta M, Mata-Cárdenas BD et al. Trichomonas vaginalis acidic phospholipase A<inf>2</inf>: Isolation and partial amino acid sequence. Acta Parasitologica. 2013 Dec 23;519-526. https://doi.org/10.2478/s11686-013-0166-2