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Abstract:

Phosphoenolpyruvate carboxykinase (PEPCK) has been purified to homogeneity from epimastigotes of the Tul 0 strain of Trypanosoma cruzi. The physicochemical parameters determined allowed the calculation of an average molecular mass of 120 kDa; the subunit molecular mass, about 61 kDa, is in good agreement with the value of 58.6 kDa recently determined from the sequence by Sommer et al. (FEBS Lett. 359 (1994) 125-129). The PEPCK from T. cruzi presented, in addition to its molecular mass, typical properties of other ATP-linked PEPCKs, namely strict specificity for ADP in the carboxylation reaction and lower specificity in the decarboxylation and exchange reactions, and synergistic activation by CdCl2 or MgCl2 when added in addition to MnCl2. The enzyme presented hysteretic behaviour, shown by a lag period in the carboxylation reaction, which was affected by dilution and preincubation. The decarboxylation reaction catalyzed by the T. cruzi PEPCK was not inhibited by excess of ATP-Mn. The apparent Km values for the carboxylation reaction, including the low value for PEP (0.035 mM) are compatible with an important role of PEPCK, as suggested by previous NMR experiments, on the CO2 fixation in vivo which leads to succinate excretion during aerobic fermentation of glucose. © 1995.

Registro:

Documento: Artículo
Título:Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
Autor:Cymeryng, C.; Cazzulo, J.J.; Cannata, J.J.B.
Filiación:Cátedra de Química Biológica, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, 1121 Buenos Aires, Argentina
Instituto de Investigaciones Bioquímicas Luis F. Leloir, Fundación Campomar - CONICET - Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, A. Machado 151, 1405 Buenos Aires, Argentina
Palabras clave:Aerobic fermentation; Glucose; Phosphoenolpyruvate carboxykinase; Trypanosoma cruzi; glucose; phosphoenolpyruvate carboxykinase (gtp); animal experiment; article; carboxylation; controlled study; enzyme analysis; enzyme purification; fermentation; nonhuman; physical chemistry; priority journal; trypanosoma cruzi; Animal; Cations, Divalent; Chemistry, Physical; Enzyme Inhibitors; Hydrogen-Ion Concentration; Kinetics; Molecular Weight; NAD; Nucleotides; Phosphoenolpyruvate Carboxykinase (GTP); Protein Conformation; Substrate Specificity; Support, Non-U.S. Gov't; Trypanosoma cruzi; Animalia; Trypanosoma; Trypanosoma cruzi
Año:1995
Volumen:73
Número:1-2
Página de inicio:91
Página de fin:101
DOI: http://dx.doi.org/10.1016/0166-6851(95)00099-M
Título revista:Molecular and Biochemical Parasitology
Título revista abreviado:Mol. Biochem. Parasitol.
ISSN:01666851
CODEN:MBIPD
CAS:Cations, Divalent; Enzyme Inhibitors; NAD, 53-84-9; Nucleotides; Phosphoenolpyruvate Carboxykinase (GTP), EC 4.1.1.32
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v73_n1-2_p91_Cymeryng

Referencias:

  • Cazzulo, The aerobic fermentation of glucose by Trypanosomatids (1992) FASEB J., 6, pp. 3153-3161
  • Bowman, Tobie, von Brand, CO2 fixation studies with the culture form of Trypanosoma cruzi (1963) Comp. Biochem. Physiol., 9, pp. 105-114
  • Cataldi de Flombaum, Cannata, Cazzulo, Segura, CO2-fixing enzymes in Trypanosoma cruzi (1977) Comp. Biochem. Physiol., 58 B, pp. 67-69
  • De los Santos, Buldain, Frydman, Cannata, Cazzulo, Carbon-13 nuclear magnetic resonance analysis of (1-13C)-glucose metabolism in Crithidia fasciculata. Evidence of CO2 fixation by phosphoenolpyruvate carboxykinase (1985) Eur. J. Biochem., 149, pp. 421-429
  • Frydman, de los Santos, Cannata, Cazzulo, Carbon-13 nuclear magnetic resonance analysis of (1-13C)glucose metabolism in Trypanosoma cruzi. Evidence of the presence of two alanine pools and of two CO2 fixation reactions (1990) Eur. J. Biochem., 192, pp. 363-368
  • Urbina, Osorno, Rojas, Inhibition of phospho enol pyruvate carboxykinase from Trypanosoma (Schyzotrypanum) cruzi epimastigotes by 3-mercaptopicolinic acid: in vitro and in vivo studies (1990) Arch. Biochem. Biophys., 282, pp. 91-99
  • Utter, Kolenbrander, Formation of oxaloacetate by CO2 fixation on phosphoenolpyruvate (1972) The Enzymes, 6, pp. 117-168. , P.D. Boyer, 2nd Edn., Academic Press, New York, NY
  • Urbina, The phosphoenol pyruvate carboxykinase of Trypanosoma (Schizotrypanum) cruzi epimastigotes: Molecular, kinetic, and regulatory properties (1987) Arch. Biochem. Biophys., 258, pp. 186-195
  • Urbina, Intermediary metabolism of Trypanosoma cruzi (1994) Parasitol. Today, 10, pp. 107-110
  • Mottram, Coombs, Purification of particulate malate dehydrogenase and phosphoenol pyruvate carboxykinase from Leishmania mexicana mexicana (1985) Biochem. Biophys. Acta, 827, pp. 310-319
  • Kueng, Schlaeppi, Schneider, Seebeck, A glycosomal protein (p60) which is predominantly expressed in procyclic Trypanosoma brucei (1989) J. Biol. Chem., 264, pp. 5203-5209
  • Parsons, Smith, Trypanosome glycosomal protein P60 is homologous to phosphoenolpyruvate carboxykinase (ATP) (1989) Nucleic Acids Res., 17, p. 6411
  • Linss, Goldenberg, Urbina, Amzel, Cloning and characterization of the gene encoding ATP-dependent phospho-enol-pyruvate carboxykinase in Trypanosoma cruzi: comparison of primary and predicted secondary structure with host GTP-dependent enzyme (1993) Gene, 136, pp. 69-77
  • Sommer, Nguyen, Wang, Phosphoenol pyruvate carboxykinase of Trypanosoma brucei is targeted to the glycosome by a C-terminal sequence (1994) FEBS Lett., 350, pp. 125-129
  • Cazzulo, Franke de Cazzulo, Engel, Cannata, End products and enzyme levels of aerobic glucose fermentation in Trypanosomatids (1985) Mol. Biochem. Parasitol., 16, pp. 329-343
  • Ernster, Zetterström, Lindberg, A method for the determination of tracer phosphate in biological material (1950) Acta Chemica Scandinavica, 4, pp. 942-947
  • Salvarrey, Cannata, Cazzulo, Phosphoenol pyruvate carboxykinase from the moderate halophile Vibrio costicola. Purification, physicochemical properties and effect of univalent-cation salts (1989) Biochem. J., 260, pp. 221-230
  • Neet, Ainslie, Hysteretic enzymes (1980) Methods Enzymol., 34, pp. 192-226
  • Warburg, Christian, Isolierung und Kristallisation des Gärungsferments Enolase (1941) Biochem. Z., 310, pp. 384-421
  • Bensadoun, Weinstein, Assay of proteins in the presence of interfering materials (1976) Anal. Biochem., 70, p. 241
  • Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685
  • Martin, Ames, A method for determining the sedimentation behavior of enzymes: application to protein mixtures (1961) J. Biol. Chem., 236, pp. 1372-1379
  • Ackers, Molecular exclusion and restricted diffusion processes in molecular sieve chromatography (1964) Biochemistry, 3, pp. 723-730
  • Cohn, Edsall, (1943) Proteins, aminoacids and peptides as ions and dipolar ions, pp. 370-381. , Reinhold, New York, NY
  • Andrews, The gel filtration behaviour of proteins related to their molecular weights over a wide range (1965) Biochem. J., 96, pp. 596-606
  • Weber, Osborn, The reliability of molecular weight determination by dodecylsulfate-polyacrylamide gel electrophoresis (1969) J. Biol. Chem., 244, pp. 4406-4412
  • Cazzulo, Stoppani, Effect of magnesium, manganese and adenosine triphosphate ions on pyruvate carboxylase from baker's yeast (1969) Biochem. J., 112, pp. 747-754
  • Tiselius, Hjertén, Levin, Protein chromatography on calcium phosphate columns (1956) Arch. Biochem. Biophys., 65, pp. 132-155
  • Cazzulo, Proteinases of Trypanosoma cruzi (1991) Biochemistry of Parasitic Protozoa, pp. 191-199. , G.H. Coombs, M.J. North, Taylor and Francis, London
  • Cannata, De Flombaum, Phosphoenolpyruvate carboxykinase from baker's yeast. Kinetics of phosphoenolpyruvate formation (1974) J. Biol. Chem., 249, pp. 3356-3365
  • Avilán, García, Hysteresis of cytosolic NADP-malic enzyme II from Trypanosoma cruzi (1994) Mol. Biochem. Parasitol., 65, pp. 225-232
  • Wright, Sanwal, Regulatory mechanisms involving nicotinamide adenine nucleotides as allosteric effectors. II. Control of phosphoenolpyruvate carboxykinase (1969) J. Biol. Chem., 244, pp. 1838-1845
  • Williamson, Mayor, Veloso, (1971) Regulation of gluconeogenesis. 9th Conference of the Gesellschaft für Biologische Chemie, pp. 92-102. , H.D. Soling, E.B. Williams, Academic Press, New York, NY
  • Wilkinson, Statistical estimations in enzyme kinetics (1961) Biochem. J., 80, pp. 324-332

Citas:

---------- APA ----------
Cymeryng, C., Cazzulo, J.J. & Cannata, J.J.B. (1995) . Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties. Molecular and Biochemical Parasitology, 73(1-2), 91-101.
http://dx.doi.org/10.1016/0166-6851(95)00099-M
---------- CHICAGO ----------
Cymeryng, C., Cazzulo, J.J., Cannata, J.J.B. "Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties" . Molecular and Biochemical Parasitology 73, no. 1-2 (1995) : 91-101.
http://dx.doi.org/10.1016/0166-6851(95)00099-M
---------- MLA ----------
Cymeryng, C., Cazzulo, J.J., Cannata, J.J.B. "Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties" . Molecular and Biochemical Parasitology, vol. 73, no. 1-2, 1995, pp. 91-101.
http://dx.doi.org/10.1016/0166-6851(95)00099-M
---------- VANCOUVER ----------
Cymeryng, C., Cazzulo, J.J., Cannata, J.J.B. Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties. Mol. Biochem. Parasitol. 1995;73(1-2):91-101.
http://dx.doi.org/10.1016/0166-6851(95)00099-M