000056238 001__ 56238
000056238 005__ 20210121114543.0
000056238 0247_ $$2doi$$a10.1074/jbc.M114.574525
000056238 0248_ $$2sideral$$a89726
000056238 037__ $$aART-2015-89726
000056238 041__ $$aeng
000056238 100__ $$aMantilla, B.S.
000056238 245__ $$aRole of ¿1-pyrroline-5-carboxylate dehydrogenase supports mitochondrial metabolism and host-cell invasion of Trypanosoma cruzi
000056238 260__ $$c2015
000056238 5060_ $$aAccess copy available to the general public$$fUnrestricted
000056238 5203_ $$aProline is crucial for energizing critical events throughout the life cycle of Trypanosoma cruzi, the etiological agent of Chagas disease. The proline breakdown pathway consists of two oxidation steps, both of which produce reducing equivalents as follows: the conversion of proline to ¿1-pyrroline-5-carboxylate (P5C), and the subsequent conversion of P5C to glutamate. We have identified and characterized the ¿1-pyrroline-5-carboxy- late dehydrogenase from T. cruzi (TcP5CDH) and report here on how this enzyme contributes to a central metabolic pathway in this parasite. Size-exclusion chromatography, two-dimen- sional gel electrophoresis, and small angle x-ray scattering analysis of TcP5CDH revealed an oligomeric state composed of two subunits of six protomers. TcP5CDH was found to complement a yeast strain deficient in PUT2 activity, confirming the enzyme’s functional role; and the biochemical parameters (Km, kcat, and kcat/Km) of the recombinant TcP5CDH were determined, exhibiting values comparable with those from T. cruzi lysates. In addition, TcP5CDH exhibited mitochondrial staining during the main stages of the T. cruzi life cycle. mRNA and enzymatic activity levels indicated the up-regulation (6-fold change) of TcP5CDH during the infective stages of the parasite. The participation of P5C as an energy source was also demonstrated. Overall, we propose that this enzymatic step is crucial for the viability of both replicative and infective forms of T. cruzi.
000056238 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000056238 590__ $$a4.258$$b2015
000056238 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b71 / 289 = 0.246$$c2015$$dQ1$$eT1
000056238 592__ $$a3.126$$b2015
000056238 593__ $$aBiochemistry$$c2015$$dQ1
000056238 593__ $$aMolecular Biology$$c2015$$dQ1
000056238 593__ $$aCell Biology$$c2015$$dQ1
000056238 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000056238 700__ $$aPaes, L.S.
000056238 700__ $$aPral, E.M.F.
000056238 700__ $$aMartil, D.E.
000056238 700__ $$aThiemann, O.H.
000056238 700__ $$0(orcid)0000-0001-8971-7355$$aFernández-Silva, P.$$uUniversidad de Zaragoza
000056238 700__ $$aBastos, E.L.
000056238 700__ $$aSilber, A.M.
000056238 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000056238 773__ $$g290, 12 (2015), 7767-7790$$pJ. biol. chem.$$tJournal of Biological Chemistry$$x0021-9258
000056238 8564_ $$s3974910$$uhttps://zaguan.unizar.es/record/56238/files/texto_completo.pdf$$yVersión publicada
000056238 8564_ $$s143424$$uhttps://zaguan.unizar.es/record/56238/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000056238 909CO $$ooai:zaguan.unizar.es:56238$$particulos$$pdriver
000056238 951__ $$a2021-01-21-11:17:42
000056238 980__ $$aARTICLE