Mass spectrometry characterization of trypanothione and novel peptides of medical importance isolated from Acanthamoeba polyphaga
Por:
Ondarza R.N., Hernandez E., Hurtado G., Woolery M., Hernandez-Sandoval F.
Publicada:
1 ene 2013
Resumen:
This paper presents unequivocal results about the presence of trypanothione and its precursor glutathionespermidine from the opportunistic human pathogen Acanthamoeba polyphaga. They were isolated by RP-HPLC as thiolbimane derivatives and characterized using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF/TOF). Additionally RP-HPLC demonstrated that thiol-bimane compounds corresponding to cysteine and glutathione were also present in A. polyphaga. Besides trypanothione, we want to report four new peptides in trophozoites, a tetrapeptide, a hexapeptide, a heptapeptide and a nonapeptide. Trypanothione and two of the thiol peptides, the hexapeptide and heptapeptide, are oxidized since the reduced forms increase in amount when the normal extract is treated by DTT or by electrolytic reduction that convert the oxidized forms to reduced ones. On the other hand, they disappear when the amoeba extract is treated with NEM or when the amoeba culture is treated with various inhibitors of NADPH-dependent disulfidereducing enzymes. Comparison of the thiol peptides, including trypanothione from A. polyphaga with extracts from human lymphocytes showed that they are not present in the latter. Therefore, some of the peptides here reported could be used as antigens for rapid detection of these parasites. In regard to the presence of the enzymes that synthesize and reduce trypanothione in A. polyphaga we suggest that they can be used as drug targets. © 2013 Bentham Science Publishers.
Filiaciones:
Department of Biochemistry, National Autonomous University of Mexico (UNAM), University City, 04510, Mexico
Center of Research on Infectious Diseases, National Institute of Public Health, Cuernavaca, Morelos 62508, Mexico
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California, San Diego (UCSD), La Jolla, CA 92093, United States
Center of Biological Research from the Nor West, CIBNOR, La Paz, Baja California Sur 23096, Mexico
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