Cloning and expression of trypanothione reductase from a New World Leishmania species
- PDF / 2,806,803 Bytes
- 10 Pages / 595 x 791 pts Page_size
- 93 Downloads / 202 Views
ORIGINAL PAPER
Cloning and expression of trypanothione reductase from a New World Leishmania species Denise Barçante Castro-Pinto · Marcelo Genestra · Gustavo B. Menezes · Mariana Waghabi · Antonio Gonçalves · Catarina De Nigris Del Cistia · Carlos Mauricio R. Sant’Anna · Leonor L. Leon · Leila Mendonça-Lima
Received: 13 June 2007 / Revised: 27 September 2007 / Accepted: 9 November 2007 / Published online: 5 December 2007 © Springer-Verlag 2007
Abstract Trypanothione disulWde (T[S]2), an unusual form of glutathione found in parasitic protozoa, plays a crucial role in the regulation of the intracellular thiol redox balance and in the defense against oxidative stress. Trypanothione reductase (TR) is central to the thiol metabolism in all trypanosomatids, including the human pathogens Trypanosoma cruzi, Trypanosoma brucei and Leishmania. Here we report the cloning, sequencing and expression of the TR encoding gene from L. (L.) amazonensis. Multiple protein sequence alignment of all known trypanosomatid TRs highlights the high degree of conservation and illustrates the phylogenetic relationships. A 3D homology model for L. amazonensis TR was constructed based on the
Communicated by Ercko Stackebrandt. Nucleotide sequence data reported in this paper is available in the GenBankTM database under accession number DQ530259. D. B. Castro-Pinto · M. Genestra · L. L. Leon Laboratory of Trypanosomatid Biochemistry, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, RJ, Brazil G. B. Menezes · M. Waghabi · L. Mendonça-Lima (&) Laboratory for Functional Genomics and Bioinformatics, Fiocruz. Av. Brasil 4365 Manguinhos, Rio de Janeiro, RJ 21040-900, Brazil e-mail: [email protected] A. Gonçalves Laboratory for Immunopathology, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, RJ, Brazil C. De Nigris Del Cistia · C. M. R. Sant’Anna Department of Chemistry, ICE, UFRRJ, Seropedica, Rio de Janeiro, RJ, Brazil
previously reported Crithidia fasciculata structure. The puriWed recombinant TR shows enzyme activity and in vivo expression of the native enzyme could be detected in infective promastigotes, both by Western blotting and by immunoXuorescence. Keywords Trypanothione reductase (EC 1.8.1.12) · Leishmania (L.) amazonensis · Thiol metabolism · Gene cloning · Leishmaniasis
Introduction A promising route towards development of improved therapeutic agents for diseases caused by human pathogens, such as parasitic protozoa, is the identiWcation of key diVerences between the metabolism of the host and the parasite, and the development of inhibitors of parasite-speciWc enzymes (Amssoms et al. 2002). Trypanothione, an unusual form of glutathione found in parasitic protozoa of the Kinetoplastidae family, plays a crucial role in regulating the intracellular thiol redox balance and in the defence against chemical and oxidative stress (reviewed in Müller et al. 2003). It contains two molecules of glutathione joined by a polyamine linker. Trypanothione reductase (TR; EC 1.8.1.12) is a NADPH-dependent Xavoprotein oxidoreductase essential in maintainin
Data Loading...
