Trichomonas vaginalis acidic phospholipase A^sub 2^: isolation and partial amino acid sequence

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 m...

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Bibliographic Details
Published in:Acta parasitologica 2013-12, Vol.58 (4), p.519
Main Authors: 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
Format: Article
Language:English
Subjects:
Amino acids
Asymptomatic
Chromatography
Enzymes
Mass spectrometry
Parasites
Proteins
Protozoa
Scientific imaging
Sexually transmitted diseases
STD
Vagina
Virulence
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Summary: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 A^sub 2^ (PLA^sub 2^) 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 A^sub 2^ 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-^sup 14^C-PA]-PC at pH 6.0. Proteins from the UFF30 sub-fraction were separated by affinity chromatography into two eluted fractions with detectable PLA A^sub 2^ 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-^sup 14^C-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 [mu]M of Rosenthal's inhibitor.[PUBLICATION ABSTRACT]
ISSN:1230-2821
1896-1851
DOI:10.2478/s11686-013-0166-2