Glutaredoxins play an important role in cellular functionality. A putative dithiol glutaredoxin is encoded in the genome of Trypanosoma cruzi. We cloned the gene and obtained the recombinant protein, which behaved as a typical thioltransferase. Activity was variable and dependent on the nature of reducer or oxidant agent used, or both. Epimastigote extracts exhibited similar activity, suggesting the occurrence of the protein in the parasite. Results support a redox scenario in T. cruzi, with glutaredoxin being involved mainly in reduction of glutathione disulfide as well as in deglutathionylation of target proteins. Antioxid. Redox Signal. 12, 787–792.
Get full access to this article
View all access options for this article.
References
1.
AriasDG, MarquezVE, BeccariaAJ, GuerreroSA, IglesiasAA. Purification and characterization of a glutathione reductase from Phaeodactylum tricornutum. Protist, 2009In press10.1016/j.protis.2009.06.001.
2.
BradfordMM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72:248–254. 1976.
Dalle-DonneI, RossiR, GiustariniD, ColomboR, MilzaniA. S-glutathionylation in protein redox regulation. Free Radic Biol Med, 43:883–898. 2007.
5.
DiscolaKF, de OliveiraMA, Rosa CussiolJR, MonteiroG, BarcenaJA, PorrasP, PadillaCA, GuimaraesBG, NettoLE. Structural aspects of the distinct biochemical properties of glutaredoxin 1 and glutaredoxin 2 from Saccharomyces cerevisiae. J Mol Biol, 385:889–901. 2009.
6.
EckersE, BienM, StroobantV, HerrmannJM, DeponteM. Biochemical characterization of dithiol glutaredoxin 8 from Saccharomyces cerevisiae: the catalytic redox mechanism redux (dagger)Biochemistry, 48:1410–1423. 2009.
7.
FairlambAH, CeramiA. Metabolism and functions of trypanothione in the Kinetoplastida. Annu Rev Microbiol, 46:695–729. 1992.
HillebrandH, SchmidtA, Krauth-SiegelRL. A second class of peroxidases linked to the trypanothione metabolism. J Biol Chem, 278:6809–6815. 2003.
11.
HirtRP, MullerS, EmbleyTM, CoombsGH. The diversity and evolution of thioredoxin reductase: new perspectives. Trends Parasitol, 18:302–308. 2002.
12.
HolmgrenA. Reduction of disulfides by thioredoxin: exceptional reactivity of insulin and suggested functions of thioredoxin in mechanism of hormone action. J Biol Chem, 254:9113–9119. 1979.
13.
Krauth-SiegelRL, CominiMA. Redox control in trypanosomatids, parasitic protozoa with trypanothione-based thiol metabolism. Biochim Biophys Acta, 1780:1236–1248. 2008.
14.
Krauth-SiegelRL, EndersB, HendersonGB, FairlambAH, SchirmerRH. Trypanothione reductase from Trypanosoma cruzi: purification and characterization of the crystalline enzyme. Eur J Biochem, 164:123–128. 1987.
15.
Krauth-SiegelRL, InhoffO. Parasite-specific trypanothione reductase as a drug target molecule. Parasitol Res, 90,suppl 2:S77–S85. 2003.
16.
LaemmliUK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227:680–685. 1970.
17.
Le TrantN, MeshnickSR, KitchenerK, EatonJW, CeramiA. Iron-containing superoxide dismutase from Crithidia fasciculata: purification, characterization, and similarity to leishmanial and trypanosomal enzymes. J Biol Chem, 258:125–130. 1983.
LudemannH, DormeyerM, SticherlingC, StallmannD, FollmannH, Krauth-SiegelRL. Trypanosoma brucei tryparedoxin, a thioredoxin-like protein in African trypanosomes. FEBS Lett, 431:381–385. 1998.
20.
LundbergM, JohanssonC, ChandraJ, EnokssonM, JacobssonG, LjungJ, JohanssonM, HolmgrenA. Cloning and expression of a novel human glutaredoxin (Grx2) with mitochondrial and nuclear isoforms. J Biol Chem, 276:26269–26275. 2001.
21.
MezzettiA, Di IlioC, CalafioreAM, AcetoA, MarzioL, FredericiG, CuccurulloF. Glutathione peroxidase, glutathione reductase and glutathione transferase activities in the human artery, vein and heart. J Mol Cell Cardiol, 22:935–938. 1990.
22.
MieyalJJ, StarkeDW, GravinaSA, DotheyC, ChungJS. Thioltransferase in human red blood cells: purification and properties. Biochemistry, 30:6088–6097. 1991.
23.
MullerS. Thioredoxin reductase and glutathione synthesis in Plasmodium falciparum. Redox Rep, 8:251–255. 2003.
24.
PiattoniCV, BlancatoVS, MigliettaH, IglesiasAA, GuerreroSA. On the occurrence of thioredoxin in Trypanosoma cruzi. Acta Trop, 97:151–160. 2006.
25.
RiusSP, CasatiP, IglesiasAA, Gomez-CasatiDF. Characterization of Arabidopsis lines deficient in GAPC-1, a cytosolic NAD-dependent glyceraldehyde-3-phosphate dehydrogenase. Plant Physiol, 148:1655–1667. 2008.
26.
SchleckerT, SchmidtA, DirdjajaN, VonckenF, ClaytonC, Krauth-SiegelRL. Substrate specificity, localization, and essential role of the glutathione peroxidase-type tryparedoxin peroxidases in Trypanosoma brucei. J Biol Chem, 280:14385–14394. 2005.
27.
ShentonD, PerroneG, QuinnKA, DawesIW, GrantCM. Regulation of protein S-thiolation by glutaredoxin 5 in the yeast Saccharomyces cerevisiae. J Biol Chem, 277:16853–16859. 2002.
28.
WilkinsonSR, MeyerDJ, KellyJM. Biochemical characterization of a trypanosome enzyme with glutathione-dependent peroxidase activity. Biochem J, 352:755–761. 2000.
29.
ZaffagniniM, MicheletL, MarchandC, SparlaF, DecottigniesP, Le MarechalP, Miginiac-MaslowM, NoctorG, TrostP, LemaireSD. The thioredoxin-independent isoform of chloroplastic glyceraldehyde-3-phosphate dehydrogenase is selectively regulated by glutathionylation. FEBS J, 274:212–226. 2007.
