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Analysis of the Trypanosoma cruzi cyclophilin gene family and identification of Cyclosporin A binding proteins

Published online by Cambridge University Press:  15 May 2006

M. POTENZA
Affiliation:
Instituto Nacional de Parasitología “Dr. M. Fatala Chabén”, A.N.L.I.S. Malbrán, Buenos Aires, Argentina
A. GALAT
Affiliation:
Departement d'Ingenierie et d'Etudes des Proteines, DSV/CEA, Gif-sur-Yvette Cedex, France
T. A. MINNING
Affiliation:
Department of Cellular Biology and Centre for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, USA
A. M. RUIZ
Affiliation:
Instituto Nacional de Parasitología “Dr. M. Fatala Chabén”, A.N.L.I.S. Malbrán, Buenos Aires, Argentina
R. DURAN
Affiliation:
Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay
R. L. TARLETON
Affiliation:
Department of Cellular Biology and Centre for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, USA
M. MARÍN
Affiliation:
Departamento de Bioquímica, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay
L. E. FICHERA
Affiliation:
Instituto Nacional de Parasitología “Dr. M. Fatala Chabén”, A.N.L.I.S. Malbrán, Buenos Aires, Argentina
J. BÚA
Affiliation:
Instituto Nacional de Parasitología “Dr. M. Fatala Chabén”, A.N.L.I.S. Malbrán, Buenos Aires, Argentina

Abstract

The Trypanosoma cruzi cyclophilin gene family comprises 15 paralogues whose nominal masses vary from 19 to 110 kDa, namely TcCyP19, TcCyP20, TcCyP21, TcCyP22, TcCyP24, TcCyP25, TcCyP26, TcCyP28, TcCyP29, TcCyP30, TcCyP34, TcCyP35, TcCyP40, TcCyP42 and TcCyP110. Under the conditions used, only some of the T. cruzi cyclophilin paralogue products could be isolated by affinity chromatography. The 15 paralogues were aligned with 495 cyclophilins from diverse organisms. Analyses of clusters formed by the T. cruzi cyclophilins with others encoded in various genomes revealed that 8 of them (TcCyP19, TcCyP21, TcCyP22, TcCyP24, TcCyP35, TcCyP40, TcCyP42 and TcCyP110) have orthologues in many different genomes whereas the other 7 display less-defined patterns of their sequence attributes and their classification to a specific group of cyclophilin's orthologues remains uncertain. Seven epimastigote cDNA clones encoding cyclophilin isoforms were further studied. These genes were found dispersed throughout the genome of the parasite. Amastigote and trypomastigote mRNAs encoding these 7 genes were also detected. We isolated 4 cyclosporin A-binding proteins in T. cruzi epimastigote extracts, which were identified by mass spectrometry as TcCyP19, TcCyP22, TcCyP28 and TcCyP40. Cyclosporin A-binding to these cyclophilins might be of importance to the mechanism of action of Cyclosporin A and its non-immunosuppressive analogues, whose trypanocidal effects were previously reported, and therefore, of potential interest in the chemotherapy of Chagas' disease.

Type
Research Article
Copyright
2006 Cambridge University Press

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