A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod
Trypanosoma cruzi adenylyl cyclases are encoded by a large polymorphic gene family. Although several genes have been identified in this parasite, little is known about the properties and regulation of these enzymes. Here we report the cloning and characterization of TczAC, a novel member of T. cruzi...
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2002
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v277_n38_p35025_DAngelo http://hdl.handle.net/20.500.12110/paper_00219258_v277_n38_p35025_DAngelo |
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paper:paper_00219258_v277_n38_p35025_DAngelo2023-06-08T14:43:21Z A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod Amino acids Calcium Catalysis Characterization Cloning Dimerization Enzymes Genes Yeast Dose-dependent stimulation Biochemistry 2' deoxyadenosine 3' phosphate adenosine triphosphate adenylate cyclase calmodulin inhibitor gene product paraflagellar rod protein protein TczAC protozoal protein unclassified drug zinc article binding affinity controlled study enzyme active site enzyme activity enzyme inhibition enzyme regulation gene expression genetic code inhibition kinetics molecular cloning multigene family nonhuman nucleotide sequence priority journal protein localization protein protein interaction protein structure regulatory mechanism Saccharomyces cerevisiae Trypanosoma cruzi Adenylate Cyclase Amino Acid Sequence Animals Base Sequence Calcium Catalytic Domain Cloning, Molecular Cyclic AMP Dimerization DNA Primers Enzyme Activation Molecular Sequence Data Protein Binding Protozoan Proteins Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Signal Transduction Trypanosoma cruzi Bovinae Mammalia Protozoa Saccharomyces Saccharomyces cerevisiae Trypanosoma Trypanosoma cruzi Trypanosoma cruzi adenylyl cyclases are encoded by a large polymorphic gene family. Although several genes have been identified in this parasite, little is known about the properties and regulation of these enzymes. Here we report the cloning and characterization of TczAC, a novel member of T. cruzi adenylyl cyclase family. The TczAC gene is expressed in all of the parasite life forms and encodes a 1,313-amino acid protein that can complement a Saccharomyces cerevisiae mutant deficient in adenylyl cyclase activity. The recombinant enzyme expressed in yeasts is constitutively active, has a low affinity for ATP (Km = 406 μM), and requires a divalent cation for catalysis. TczAC is inhibited by Zn2+ and the P-site inhibitor 2′-deoxyadenosine 3′-monophosphate, suggesting some level of conservation in the catalytic mechanism with mammalian adenylyl cyclases. It shows a dose-dependent stimulation by Ca2+ which can be reversed by high concentrations of phenothiazinic calmodulin inhibitors. However, bovine calmodulin fails to stimulate the enzyme. Using a yeast two-hybrid screen it was found that TczAC interacts through its catalytic domain with the paraflagellar rod protein, a component of the flagellar structure. Furthermore, we demonstrate that TczAC can dimerize through the same domain. These results provide novel evidence of the possible localization and regulation of this protein. 2002 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v277_n38_p35025_DAngelo http://hdl.handle.net/20.500.12110/paper_00219258_v277_n38_p35025_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 |
Amino acids Calcium Catalysis Characterization Cloning Dimerization Enzymes Genes Yeast Dose-dependent stimulation Biochemistry 2' deoxyadenosine 3' phosphate adenosine triphosphate adenylate cyclase calmodulin inhibitor gene product paraflagellar rod protein protein TczAC protozoal protein unclassified drug zinc article binding affinity controlled study enzyme active site enzyme activity enzyme inhibition enzyme regulation gene expression genetic code inhibition kinetics molecular cloning multigene family nonhuman nucleotide sequence priority journal protein localization protein protein interaction protein structure regulatory mechanism Saccharomyces cerevisiae Trypanosoma cruzi Adenylate Cyclase Amino Acid Sequence Animals Base Sequence Calcium Catalytic Domain Cloning, Molecular Cyclic AMP Dimerization DNA Primers Enzyme Activation Molecular Sequence Data Protein Binding Protozoan Proteins Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Signal Transduction Trypanosoma cruzi Bovinae Mammalia Protozoa Saccharomyces Saccharomyces cerevisiae Trypanosoma Trypanosoma cruzi |
| spellingShingle |
Amino acids Calcium Catalysis Characterization Cloning Dimerization Enzymes Genes Yeast Dose-dependent stimulation Biochemistry 2' deoxyadenosine 3' phosphate adenosine triphosphate adenylate cyclase calmodulin inhibitor gene product paraflagellar rod protein protein TczAC protozoal protein unclassified drug zinc article binding affinity controlled study enzyme active site enzyme activity enzyme inhibition enzyme regulation gene expression genetic code inhibition kinetics molecular cloning multigene family nonhuman nucleotide sequence priority journal protein localization protein protein interaction protein structure regulatory mechanism Saccharomyces cerevisiae Trypanosoma cruzi Adenylate Cyclase Amino Acid Sequence Animals Base Sequence Calcium Catalytic Domain Cloning, Molecular Cyclic AMP Dimerization DNA Primers Enzyme Activation Molecular Sequence Data Protein Binding Protozoan Proteins Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Signal Transduction Trypanosoma cruzi Bovinae Mammalia Protozoa Saccharomyces Saccharomyces cerevisiae Trypanosoma Trypanosoma cruzi A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| topic_facet |
Amino acids Calcium Catalysis Characterization Cloning Dimerization Enzymes Genes Yeast Dose-dependent stimulation Biochemistry 2' deoxyadenosine 3' phosphate adenosine triphosphate adenylate cyclase calmodulin inhibitor gene product paraflagellar rod protein protein TczAC protozoal protein unclassified drug zinc article binding affinity controlled study enzyme active site enzyme activity enzyme inhibition enzyme regulation gene expression genetic code inhibition kinetics molecular cloning multigene family nonhuman nucleotide sequence priority journal protein localization protein protein interaction protein structure regulatory mechanism Saccharomyces cerevisiae Trypanosoma cruzi Adenylate Cyclase Amino Acid Sequence Animals Base Sequence Calcium Catalytic Domain Cloning, Molecular Cyclic AMP Dimerization DNA Primers Enzyme Activation Molecular Sequence Data Protein Binding Protozoan Proteins Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid Signal Transduction Trypanosoma cruzi Bovinae Mammalia Protozoa Saccharomyces Saccharomyces cerevisiae Trypanosoma Trypanosoma cruzi |
| description |
Trypanosoma cruzi adenylyl cyclases are encoded by a large polymorphic gene family. Although several genes have been identified in this parasite, little is known about the properties and regulation of these enzymes. Here we report the cloning and characterization of TczAC, a novel member of T. cruzi adenylyl cyclase family. The TczAC gene is expressed in all of the parasite life forms and encodes a 1,313-amino acid protein that can complement a Saccharomyces cerevisiae mutant deficient in adenylyl cyclase activity. The recombinant enzyme expressed in yeasts is constitutively active, has a low affinity for ATP (Km = 406 μM), and requires a divalent cation for catalysis. TczAC is inhibited by Zn2+ and the P-site inhibitor 2′-deoxyadenosine 3′-monophosphate, suggesting some level of conservation in the catalytic mechanism with mammalian adenylyl cyclases. It shows a dose-dependent stimulation by Ca2+ which can be reversed by high concentrations of phenothiazinic calmodulin inhibitors. However, bovine calmodulin fails to stimulate the enzyme. Using a yeast two-hybrid screen it was found that TczAC interacts through its catalytic domain with the paraflagellar rod protein, a component of the flagellar structure. Furthermore, we demonstrate that TczAC can dimerize through the same domain. These results provide novel evidence of the possible localization and regulation of this protein. |
| title |
A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| title_short |
A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| title_full |
A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| title_fullStr |
A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| title_full_unstemmed |
A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| title_sort |
novel calcium-stimulated adenylyl cyclase from trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod |
| publishDate |
2002 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v277_n38_p35025_DAngelo http://hdl.handle.net/20.500.12110/paper_00219258_v277_n38_p35025_DAngelo |
| _version_ |
1768543166230691840 |