Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties

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 agre...

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Autores principales:	Cymeryng, Cora Beatriz, Cannata, Joaquín J.B.
Publicado:	1995
Materias:	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 Animalia Trypanosoma
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v73_n1-2_p91_Cymeryng http://hdl.handle.net/20.500.12110/paper_01666851_v73_n1-2_p91_Cymeryng
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_01666851_v73_n1-2_p91_Cymeryng
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spelling	paper:paper_01666851_v73_n1-2_p91_Cymeryng2023-06-08T15:16:07Z Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties Cymeryng, Cora Beatriz Cannata, Joaquín J.B. 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 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. Fil:Cymeryng, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cannata, J.J.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1995 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v73_n1-2_p91_Cymeryng http://hdl.handle.net/20.500.12110/paper_01666851_v73_n1-2_p91_Cymeryng
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 Cymeryng, Cora Beatriz Cannata, Joaquín J.B. Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
topic_facet	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
description	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.
author	Cymeryng, Cora Beatriz Cannata, Joaquín J.B.
author_facet	Cymeryng, Cora Beatriz Cannata, Joaquín J.B.
author_sort	Cymeryng, Cora Beatriz
title	Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
title_short	Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
title_full	Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
title_fullStr	Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
title_full_unstemmed	Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties
title_sort	phosphoenolpyruvate carboxykinase from trypanosoma cruzi. purification and physicochemical and kinetic properties
publishDate	1995
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v73_n1-2_p91_Cymeryng http://hdl.handle.net/20.500.12110/paper_01666851_v73_n1-2_p91_Cymeryng
work_keys_str_mv	AT cymeryngcorabeatriz phosphoenolpyruvatecarboxykinasefromtrypanosomacruzipurificationandphysicochemicalandkineticproperties AT cannatajoaquinjb phosphoenolpyruvatecarboxykinasefromtrypanosomacruzipurificationandphysicochemicalandkineticproperties
_version_	1768544405479751680

Phosphoenolpyruvate carboxykinase from Trypanosoma cruzi. Purification and physicochemical and kinetic properties

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