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Originally published In Press as doi:10.1074/jbc.M003893200 on June 27, 2000

J. Biol. Chem., Vol. 275, Issue 37, 28356-28362, September 15, 2000
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31P NMR Spectroscopy of Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major
EVIDENCE FOR HIGH LEVELS OF CONDENSED INORGANIC PHOSPHATES*

Benjamin Moreno§, Julio A. Urbina, Eric Oldfield||, Brian N. Bailey**Dagger Dagger , Claudia O. Rodrigues**, and Roberto Docampo**

From the Departments of Chemistry and Biophysics, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801 and the ** Laboratory of Molecular Parasitology, Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802

High resolution 31P nuclear magnetic resonance spectra at 303.6 MHz (corresponding to a 1H resonance frequency of 750 MHz) have been obtained of perchloric acid extracts of Trypanosoma brucei, Trypanosoma cruzi, and Leishmania major, the causative agents of African sleeping sickness, Chagas' disease, and leishmaniasis. Essentially complete assignments have been made based on chemical shifts and by direct addition of authentic reference compounds. The results indicate the presence of high levels of short chain condensed polyphosphates: di-, tri-, tetra-, and pentapolyphosphate. 31P NMR spectra of purified T. brucei, T. cruzi, and L. major acidocalcisomes, calcium and phosphorus storage organelles, indicate that polyphosphates are abundant in these organelles and have an average chain length of 3.11-3.39 phosphates. In the context of the recent discovery of several pyrophosphate-utilizing enzymes in trypanosomatids, the presence of these inorganic polyphosphates implies a critical role for these molecules in these parasites and a potential new route to chemotherapy.

* This work was supported in part by National Institutes of Health Grants GM-50694 (to E. O.) and AI-23259 (to R. D.) and by Grants from the United Nations Development Programme/World Bank/World Health Organization Special Programme for Research and Training in Tropical Diseases (to R. D. and J. A. U.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by an Illinois Minority Graduate Incentive Program fellowship.

Permanent address: Instituto Venezolano de Investigaciones Científicas, Caracas, 1020 Venezuela.

|| To whom correspondence should be addressed: Depts. of Chemistry and Biophysics, University of Illinois at Urbana-Champaign, 600 S. Mathews Ave., Urbana, IL 61801. Fax: 217-244-0997; E-mail: eo@chad. scs.uiuc.edu.

Dagger Dagger Supported by a student fellowship from the American Heart Association, Midwest Affiliate.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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