Regulation of Ca2+/calmodulin-dependent protein kinase from
A multifunctional Ca2+/calmodulin-dependent protein kinase (TcCaM K) was purified and characterized from the cytosol of Trypanosoma cruzi epimastigote forms. Like mammalian CaM KII, TcCaM K has a broad substrate specificity and a similar subunit composition. Western blot analysis revealed that this...
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todo:paper_01666851_v78_n1-2_p171_Cruzi2023-10-03T15:04:24Z Regulation of Ca2+/calmodulin-dependent protein kinase from Cruzi, T. Ogueta, S. Intosh, G.M. Téllez-Iñon, Ma.T. /cam-dependent protein kinase Ca2+ Regulation Trypanosoma cruzi Mammalia Trypanosoma Trypanosoma cruzi protein kinase (calcium,calmodulin) monoclonal antibody peptide protein kinase (calcium,calmodulin) protozoon antibody syntide 2 syntide-2 article autophosphorylation calcium transport enzyme purification enzyme regulation enzyme specificity enzyme subunit membrane binding priority journal trypanosoma cruzi amino acid sequence animal biological model chemistry enzymology immunology isolation and purification kinetics metabolism molecular genetics molecular weight phosphorylation protein conformation rat solubility Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Antibodies, Protozoan Ca(2+)-Calmodulin Dependent Protein Kinase Kinetics Models, Biological Molecular Sequence Data Molecular Weight Peptides Phosphorylation Protein Conformation Rats Solubility Substrate Specificity Trypanosoma cruzi A multifunctional Ca2+/calmodulin-dependent protein kinase (TcCaM K) was purified and characterized from the cytosol of Trypanosoma cruzi epimastigote forms. Like mammalian CaM KII, TcCaM K has a broad substrate specificity and a similar subunit composition. Western blot analysis revealed that this TcCaM K possesses two subunits of 50 and 60 kDa, which exhibited autophosphorylating activity. A panel of monoclonal and polyclonal antibodies raised against rat brain CaM KII could also recognize TcCaM K. However, experimental evidence suggests a different conformational arrangement of the TcCaM K subunits. Like its mammalian counterpart, two highly active autonomous, Ca2+-independent, states of TcCaM K can be isolated. These states, caused by high phosphate incorporation, differ only in their extent of Ca2+/CaM-dependence. About 15-20% of the autophosphorylated TcCaM K can be reverted using protein phosphatase 2A, and, consequently, its Ca2+-dependent activity is also partially restored. The situation is somewhat different when the enzyme is linked to the cytoskeleton, as we have previously shown. The membrane-bound form is present only in the native form. Activation increases its protein kinase activity from 5-to 14-fold. In this study, we provide evidence of another form of TcCaM K present in soluble fractions of the parasite that can be isolated in autonomous states. Our results suggest that autophosphorylation of membrane-bound TcCaM K may be responsible for kinase release in a Ca2+/CaM-independent state. These properties of TcCaM K may play an important role in regulating Ca2+-dependent processes in the parasite. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_01666851_v78_n1-2_p171_Cruzi |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
/cam-dependent protein kinase Ca2+ Regulation Trypanosoma cruzi Mammalia Trypanosoma Trypanosoma cruzi protein kinase (calcium,calmodulin) monoclonal antibody peptide protein kinase (calcium,calmodulin) protozoon antibody syntide 2 syntide-2 article autophosphorylation calcium transport enzyme purification enzyme regulation enzyme specificity enzyme subunit membrane binding priority journal trypanosoma cruzi amino acid sequence animal biological model chemistry enzymology immunology isolation and purification kinetics metabolism molecular genetics molecular weight phosphorylation protein conformation rat solubility Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Antibodies, Protozoan Ca(2+)-Calmodulin Dependent Protein Kinase Kinetics Models, Biological Molecular Sequence Data Molecular Weight Peptides Phosphorylation Protein Conformation Rats Solubility Substrate Specificity Trypanosoma cruzi |
| spellingShingle |
/cam-dependent protein kinase Ca2+ Regulation Trypanosoma cruzi Mammalia Trypanosoma Trypanosoma cruzi protein kinase (calcium,calmodulin) monoclonal antibody peptide protein kinase (calcium,calmodulin) protozoon antibody syntide 2 syntide-2 article autophosphorylation calcium transport enzyme purification enzyme regulation enzyme specificity enzyme subunit membrane binding priority journal trypanosoma cruzi amino acid sequence animal biological model chemistry enzymology immunology isolation and purification kinetics metabolism molecular genetics molecular weight phosphorylation protein conformation rat solubility Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Antibodies, Protozoan Ca(2+)-Calmodulin Dependent Protein Kinase Kinetics Models, Biological Molecular Sequence Data Molecular Weight Peptides Phosphorylation Protein Conformation Rats Solubility Substrate Specificity Trypanosoma cruzi Cruzi, T. Ogueta, S. Intosh, G.M. Téllez-Iñon, Ma.T. Regulation of Ca2+/calmodulin-dependent protein kinase from |
| topic_facet |
/cam-dependent protein kinase Ca2+ Regulation Trypanosoma cruzi Mammalia Trypanosoma Trypanosoma cruzi protein kinase (calcium,calmodulin) monoclonal antibody peptide protein kinase (calcium,calmodulin) protozoon antibody syntide 2 syntide-2 article autophosphorylation calcium transport enzyme purification enzyme regulation enzyme specificity enzyme subunit membrane binding priority journal trypanosoma cruzi amino acid sequence animal biological model chemistry enzymology immunology isolation and purification kinetics metabolism molecular genetics molecular weight phosphorylation protein conformation rat solubility Trypanosoma cruzi Amino Acid Sequence Animals Antibodies, Monoclonal Antibodies, Protozoan Ca(2+)-Calmodulin Dependent Protein Kinase Kinetics Models, Biological Molecular Sequence Data Molecular Weight Peptides Phosphorylation Protein Conformation Rats Solubility Substrate Specificity Trypanosoma cruzi |
| description |
A multifunctional Ca2+/calmodulin-dependent protein kinase (TcCaM K) was purified and characterized from the cytosol of Trypanosoma cruzi epimastigote forms. Like mammalian CaM KII, TcCaM K has a broad substrate specificity and a similar subunit composition. Western blot analysis revealed that this TcCaM K possesses two subunits of 50 and 60 kDa, which exhibited autophosphorylating activity. A panel of monoclonal and polyclonal antibodies raised against rat brain CaM KII could also recognize TcCaM K. However, experimental evidence suggests a different conformational arrangement of the TcCaM K subunits. Like its mammalian counterpart, two highly active autonomous, Ca2+-independent, states of TcCaM K can be isolated. These states, caused by high phosphate incorporation, differ only in their extent of Ca2+/CaM-dependence. About 15-20% of the autophosphorylated TcCaM K can be reverted using protein phosphatase 2A, and, consequently, its Ca2+-dependent activity is also partially restored. The situation is somewhat different when the enzyme is linked to the cytoskeleton, as we have previously shown. The membrane-bound form is present only in the native form. Activation increases its protein kinase activity from 5-to 14-fold. In this study, we provide evidence of another form of TcCaM K present in soluble fractions of the parasite that can be isolated in autonomous states. Our results suggest that autophosphorylation of membrane-bound TcCaM K may be responsible for kinase release in a Ca2+/CaM-independent state. These properties of TcCaM K may play an important role in regulating Ca2+-dependent processes in the parasite. |
| format |
JOUR |
| author |
Cruzi, T. Ogueta, S. Intosh, G.M. Téllez-Iñon, Ma.T. |
| author_facet |
Cruzi, T. Ogueta, S. Intosh, G.M. Téllez-Iñon, Ma.T. |
| author_sort |
Cruzi, T. |
| title |
Regulation of Ca2+/calmodulin-dependent protein kinase from |
| title_short |
Regulation of Ca2+/calmodulin-dependent protein kinase from |
| title_full |
Regulation of Ca2+/calmodulin-dependent protein kinase from |
| title_fullStr |
Regulation of Ca2+/calmodulin-dependent protein kinase from |
| title_full_unstemmed |
Regulation of Ca2+/calmodulin-dependent protein kinase from |
| title_sort |
regulation of ca2+/calmodulin-dependent protein kinase from |
| url |
http://hdl.handle.net/20.500.12110/paper_01666851_v78_n1-2_p171_Cruzi |
| work_keys_str_mv |
AT cruzit regulationofca2calmodulindependentproteinkinasefrom AT oguetas regulationofca2calmodulindependentproteinkinasefrom AT intoshgm regulationofca2calmodulindependentproteinkinasefrom AT tellezinonmat regulationofca2calmodulindependentproteinkinasefrom |
| _version_ |
1807323171318136832 |