Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi
Compartmentalization of cAMP phosphodiesterases plays a key role in the regulation of cAMP signalling in mammals. In the present paper, we report the characterization and subcellular localization of TcPDE1, the first cAMP-specific phosphodiesterase to be identified from Trypanosoma cruzi. TcPDE1 is...
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todo:paper_02646021_v378_n1_p63_DAngelo2023-10-03T15:13:00Z Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi D'Angelo, M.A. Sanguineti, S. Reece, J.M. Birnbaumer, L. Torres, H.N. Flawiá, M.M. cAMP Flagellum Membrane Phosphodiesterase Trypanosoma cruzi Biological membranes Enzyme inhibition Enzymes Genes Hydrochloric acid Yeast Cell disruption Mutants Biochemistry 9 (2 hydroxy 3 nonyl)adenine amino acid calcium channel blocking agent calmodulin inhibitor chloride cyclic AMP cyclic AMP phosphodiesterase cyclic AMP phosphodiesterase 1 cyclic GMP dipyridamole enzyme isobutylmethylxanthine membrane protein papaverine phosphodiesterase phosphodiesterase inhibitor protein inhibitor protein subunit recombinant protein rolipram sodium chloride theophylline unclassified drug vinpocetine zaprinast article cell disruption cell fractionation cell membrane cell organelle cellular distribution chemosensitivity confocal laser microscopy controlled study enzyme activity enzyme localization enzyme specificity epimastigote flagellum genetic code heat shock membrane multigene family nonhuman nucleotide sequence parasite priority journal protein analysis protein deficiency protein secretion temperature sensitive mutant Trypanosoma Trypanosoma cruzi yeast 3',5'-Cyclic-Nucleotide Phosphodiesterase Amino Acid Sequence Animals Cell Membrane Cloning, Molecular Flagella Gene Components Genetic Complementation Test Microscopy, Confocal Molecular Sequence Data Subcellular Fractions Trypanosoma cruzi Yeasts Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae Compartmentalization of cAMP phosphodiesterases plays a key role in the regulation of cAMP signalling in mammals. In the present paper, we report the characterization and subcellular localization of TcPDE1, the first cAMP-specific phosphodiesterase to be identified from Trypanosoma cruzi. TcPDE1 is part of a small gene family and encodes a 929-amino-acid protein that can complement a heat-shock-sensitive yeast mutant deficient in phosphodiesterase genes. Recombinant TcPDE1 strongly associates with membranes and cannot be released with NaCl or sodium cholate, suggesting that it is an integral membrane protein. This enzyme is specific for cAMP and its activity is not affected by cGMP, Ca2+, calmodulin or fenotiazinic inhibitors. TcPDE1 is sensitive to the phosphodiesterase inhibitor dipyridamole but is resistant to 3-isobutyl-1-methylxanthine, theophylline, rolipram and zaprinast. Papaverine, erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride, and vinpocetine are poor inhibitors of this enzyme. Confocal laser scanning of T. cruzi epimastigotes showed that TcPDE1 is associated with the plasma membrane and concentrated in the flagellum of the parasite. The association of TcPDE1 with this organelle was confirmed by subcellular fractionation and cell-disruption treatments. The localization of this enzyme is a unique feature that distinguishes it from all the trypanosomatid phosphodiesterases described so far and indicates that compartmentalization of cAMP phosphodiesterases could also be important in these parasites. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_02646021_v378_n1_p63_DAngelo |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
cAMP Flagellum Membrane Phosphodiesterase Trypanosoma cruzi Biological membranes Enzyme inhibition Enzymes Genes Hydrochloric acid Yeast Cell disruption Mutants Biochemistry 9 (2 hydroxy 3 nonyl)adenine amino acid calcium channel blocking agent calmodulin inhibitor chloride cyclic AMP cyclic AMP phosphodiesterase cyclic AMP phosphodiesterase 1 cyclic GMP dipyridamole enzyme isobutylmethylxanthine membrane protein papaverine phosphodiesterase phosphodiesterase inhibitor protein inhibitor protein subunit recombinant protein rolipram sodium chloride theophylline unclassified drug vinpocetine zaprinast article cell disruption cell fractionation cell membrane cell organelle cellular distribution chemosensitivity confocal laser microscopy controlled study enzyme activity enzyme localization enzyme specificity epimastigote flagellum genetic code heat shock membrane multigene family nonhuman nucleotide sequence parasite priority journal protein analysis protein deficiency protein secretion temperature sensitive mutant Trypanosoma Trypanosoma cruzi yeast 3',5'-Cyclic-Nucleotide Phosphodiesterase Amino Acid Sequence Animals Cell Membrane Cloning, Molecular Flagella Gene Components Genetic Complementation Test Microscopy, Confocal Molecular Sequence Data Subcellular Fractions Trypanosoma cruzi Yeasts Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae |
| spellingShingle |
cAMP Flagellum Membrane Phosphodiesterase Trypanosoma cruzi Biological membranes Enzyme inhibition Enzymes Genes Hydrochloric acid Yeast Cell disruption Mutants Biochemistry 9 (2 hydroxy 3 nonyl)adenine amino acid calcium channel blocking agent calmodulin inhibitor chloride cyclic AMP cyclic AMP phosphodiesterase cyclic AMP phosphodiesterase 1 cyclic GMP dipyridamole enzyme isobutylmethylxanthine membrane protein papaverine phosphodiesterase phosphodiesterase inhibitor protein inhibitor protein subunit recombinant protein rolipram sodium chloride theophylline unclassified drug vinpocetine zaprinast article cell disruption cell fractionation cell membrane cell organelle cellular distribution chemosensitivity confocal laser microscopy controlled study enzyme activity enzyme localization enzyme specificity epimastigote flagellum genetic code heat shock membrane multigene family nonhuman nucleotide sequence parasite priority journal protein analysis protein deficiency protein secretion temperature sensitive mutant Trypanosoma Trypanosoma cruzi yeast 3',5'-Cyclic-Nucleotide Phosphodiesterase Amino Acid Sequence Animals Cell Membrane Cloning, Molecular Flagella Gene Components Genetic Complementation Test Microscopy, Confocal Molecular Sequence Data Subcellular Fractions Trypanosoma cruzi Yeasts Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae D'Angelo, M.A. Sanguineti, S. Reece, J.M. Birnbaumer, L. Torres, H.N. Flawiá, M.M. Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| topic_facet |
cAMP Flagellum Membrane Phosphodiesterase Trypanosoma cruzi Biological membranes Enzyme inhibition Enzymes Genes Hydrochloric acid Yeast Cell disruption Mutants Biochemistry 9 (2 hydroxy 3 nonyl)adenine amino acid calcium channel blocking agent calmodulin inhibitor chloride cyclic AMP cyclic AMP phosphodiesterase cyclic AMP phosphodiesterase 1 cyclic GMP dipyridamole enzyme isobutylmethylxanthine membrane protein papaverine phosphodiesterase phosphodiesterase inhibitor protein inhibitor protein subunit recombinant protein rolipram sodium chloride theophylline unclassified drug vinpocetine zaprinast article cell disruption cell fractionation cell membrane cell organelle cellular distribution chemosensitivity confocal laser microscopy controlled study enzyme activity enzyme localization enzyme specificity epimastigote flagellum genetic code heat shock membrane multigene family nonhuman nucleotide sequence parasite priority journal protein analysis protein deficiency protein secretion temperature sensitive mutant Trypanosoma Trypanosoma cruzi yeast 3',5'-Cyclic-Nucleotide Phosphodiesterase Amino Acid Sequence Animals Cell Membrane Cloning, Molecular Flagella Gene Components Genetic Complementation Test Microscopy, Confocal Molecular Sequence Data Subcellular Fractions Trypanosoma cruzi Yeasts Mammalia Protozoa Trypanosoma Trypanosoma cruzi Trypanosomatidae |
| description |
Compartmentalization of cAMP phosphodiesterases plays a key role in the regulation of cAMP signalling in mammals. In the present paper, we report the characterization and subcellular localization of TcPDE1, the first cAMP-specific phosphodiesterase to be identified from Trypanosoma cruzi. TcPDE1 is part of a small gene family and encodes a 929-amino-acid protein that can complement a heat-shock-sensitive yeast mutant deficient in phosphodiesterase genes. Recombinant TcPDE1 strongly associates with membranes and cannot be released with NaCl or sodium cholate, suggesting that it is an integral membrane protein. This enzyme is specific for cAMP and its activity is not affected by cGMP, Ca2+, calmodulin or fenotiazinic inhibitors. TcPDE1 is sensitive to the phosphodiesterase inhibitor dipyridamole but is resistant to 3-isobutyl-1-methylxanthine, theophylline, rolipram and zaprinast. Papaverine, erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride, and vinpocetine are poor inhibitors of this enzyme. Confocal laser scanning of T. cruzi epimastigotes showed that TcPDE1 is associated with the plasma membrane and concentrated in the flagellum of the parasite. The association of TcPDE1 with this organelle was confirmed by subcellular fractionation and cell-disruption treatments. The localization of this enzyme is a unique feature that distinguishes it from all the trypanosomatid phosphodiesterases described so far and indicates that compartmentalization of cAMP phosphodiesterases could also be important in these parasites. |
| format |
JOUR |
| author |
D'Angelo, M.A. Sanguineti, S. Reece, J.M. Birnbaumer, L. Torres, H.N. Flawiá, M.M. |
| author_facet |
D'Angelo, M.A. Sanguineti, S. Reece, J.M. Birnbaumer, L. Torres, H.N. Flawiá, M.M. |
| author_sort |
D'Angelo, M.A. |
| title |
Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| title_short |
Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| title_full |
Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| title_fullStr |
Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| title_full_unstemmed |
Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi |
| title_sort |
identification, characterization and subcellular localization of tcpde1, a novel camp-specific phosphodiesterase from trypanosoma cruzi |
| url |
http://hdl.handle.net/20.500.12110/paper_02646021_v378_n1_p63_DAngelo |
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