A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity
The strength of the interaction between the catalytic and regulatory subunits in protein kinase A differs among species. The linker region from regulatory subunits is non-conserved. To evaluate the participation of this region in the interaction with the catalytic subunit, we have assayed its effect...
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2008
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00039861_v480_n2_p95_Rinaldi http://hdl.handle.net/20.500.12110/paper_00039861_v480_n2_p95_Rinaldi |
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paper:paper_00039861_v480_n2_p95_Rinaldi2023-06-08T14:25:05Z A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity Rinaldi, Jimena Julieta Ocampo, Josefina Rossi, Silvia Graciela Moreno de Colonna, Silvia Activation Inhibition Linker domain I Peptides Protein kinase A Regulatory subunit Substrates cyclic AMP dependent protein kinase kemptide synthetic peptide amino terminal sequence article autophosphorylation competitive inhibition enzyme activation enzyme active site enzyme activity enzyme kinetics enzyme substrate enzyme subunit molecular interaction Mucor mucor circinelloides mucor rouxii nonhuman priority journal protein domain protein phosphorylation Saccharomyces cerevisiae sequence homology species difference viscosity Amino Acid Sequence Amino Acids Catalysis Catalytic Domain Cyclic AMP-Dependent Protein Kinases Enzyme Activation Gene Expression Regulation, Fungal Kinetics Molecular Sequence Data Mucor Peptides Phosphorylation Protein Structure, Tertiary Saccharomyces cerevisiae Sequence Homology, Amino Acid Amylomyces rouxii Fungi Mucor circinelloides Saccharomyces cerevisiae The strength of the interaction between the catalytic and regulatory subunits in protein kinase A differs among species. The linker region from regulatory subunits is non-conserved. To evaluate the participation of this region in the interaction with the catalytic subunit, we have assayed its effect on the enzymatic properties of the catalytic subunit. Protein kinase A from three fungi, Mucor rouxii, Mucor circinelloides and Saccharomyces cerevisiae have been chosen as models. The R-C interaction is explored by using synthetic peptides of 8, 18 and 47 amino acids, corresponding to the R subunit autophosphorylation site plus a variable region toward the N terminus (0, 10, or 39 residues). The Km of the catalytic subunits decreased with the length of the peptide, while the Vmax increased. Viscosity studies identified product release as the rate limiting step for phosphorylation of the longer peptides. Pseudosubstrate derivatives of the 18 residue peptides did not display a competitive inhibition behavior toward either kemptide or a bona fide protein substrate since, at low relative pseudosubstrate/substrate concentration, stimulation of kemptide or protein substrate phosphorylation was observed. The behavior was mimicked by intact R. We conclude that in addition to its negative regulatory role, the R subunit stimulates C activity via distal interactions. © 2008 Elsevier Inc. All rights reserved. Fil:Rinaldi, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ocampo, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Rossi, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moreno, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2008 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00039861_v480_n2_p95_Rinaldi http://hdl.handle.net/20.500.12110/paper_00039861_v480_n2_p95_Rinaldi |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Activation Inhibition Linker domain I Peptides Protein kinase A Regulatory subunit Substrates cyclic AMP dependent protein kinase kemptide synthetic peptide amino terminal sequence article autophosphorylation competitive inhibition enzyme activation enzyme active site enzyme activity enzyme kinetics enzyme substrate enzyme subunit molecular interaction Mucor mucor circinelloides mucor rouxii nonhuman priority journal protein domain protein phosphorylation Saccharomyces cerevisiae sequence homology species difference viscosity Amino Acid Sequence Amino Acids Catalysis Catalytic Domain Cyclic AMP-Dependent Protein Kinases Enzyme Activation Gene Expression Regulation, Fungal Kinetics Molecular Sequence Data Mucor Peptides Phosphorylation Protein Structure, Tertiary Saccharomyces cerevisiae Sequence Homology, Amino Acid Amylomyces rouxii Fungi Mucor circinelloides Saccharomyces cerevisiae |
| spellingShingle |
Activation Inhibition Linker domain I Peptides Protein kinase A Regulatory subunit Substrates cyclic AMP dependent protein kinase kemptide synthetic peptide amino terminal sequence article autophosphorylation competitive inhibition enzyme activation enzyme active site enzyme activity enzyme kinetics enzyme substrate enzyme subunit molecular interaction Mucor mucor circinelloides mucor rouxii nonhuman priority journal protein domain protein phosphorylation Saccharomyces cerevisiae sequence homology species difference viscosity Amino Acid Sequence Amino Acids Catalysis Catalytic Domain Cyclic AMP-Dependent Protein Kinases Enzyme Activation Gene Expression Regulation, Fungal Kinetics Molecular Sequence Data Mucor Peptides Phosphorylation Protein Structure, Tertiary Saccharomyces cerevisiae Sequence Homology, Amino Acid Amylomyces rouxii Fungi Mucor circinelloides Saccharomyces cerevisiae Rinaldi, Jimena Julieta Ocampo, Josefina Rossi, Silvia Graciela Moreno de Colonna, Silvia A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| topic_facet |
Activation Inhibition Linker domain I Peptides Protein kinase A Regulatory subunit Substrates cyclic AMP dependent protein kinase kemptide synthetic peptide amino terminal sequence article autophosphorylation competitive inhibition enzyme activation enzyme active site enzyme activity enzyme kinetics enzyme substrate enzyme subunit molecular interaction Mucor mucor circinelloides mucor rouxii nonhuman priority journal protein domain protein phosphorylation Saccharomyces cerevisiae sequence homology species difference viscosity Amino Acid Sequence Amino Acids Catalysis Catalytic Domain Cyclic AMP-Dependent Protein Kinases Enzyme Activation Gene Expression Regulation, Fungal Kinetics Molecular Sequence Data Mucor Peptides Phosphorylation Protein Structure, Tertiary Saccharomyces cerevisiae Sequence Homology, Amino Acid Amylomyces rouxii Fungi Mucor circinelloides Saccharomyces cerevisiae |
| description |
The strength of the interaction between the catalytic and regulatory subunits in protein kinase A differs among species. The linker region from regulatory subunits is non-conserved. To evaluate the participation of this region in the interaction with the catalytic subunit, we have assayed its effect on the enzymatic properties of the catalytic subunit. Protein kinase A from three fungi, Mucor rouxii, Mucor circinelloides and Saccharomyces cerevisiae have been chosen as models. The R-C interaction is explored by using synthetic peptides of 8, 18 and 47 amino acids, corresponding to the R subunit autophosphorylation site plus a variable region toward the N terminus (0, 10, or 39 residues). The Km of the catalytic subunits decreased with the length of the peptide, while the Vmax increased. Viscosity studies identified product release as the rate limiting step for phosphorylation of the longer peptides. Pseudosubstrate derivatives of the 18 residue peptides did not display a competitive inhibition behavior toward either kemptide or a bona fide protein substrate since, at low relative pseudosubstrate/substrate concentration, stimulation of kemptide or protein substrate phosphorylation was observed. The behavior was mimicked by intact R. We conclude that in addition to its negative regulatory role, the R subunit stimulates C activity via distal interactions. © 2008 Elsevier Inc. All rights reserved. |
| author |
Rinaldi, Jimena Julieta Ocampo, Josefina Rossi, Silvia Graciela Moreno de Colonna, Silvia |
| author_facet |
Rinaldi, Jimena Julieta Ocampo, Josefina Rossi, Silvia Graciela Moreno de Colonna, Silvia |
| author_sort |
Rinaldi, Jimena Julieta |
| title |
A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| title_short |
A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| title_full |
A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| title_fullStr |
A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| title_full_unstemmed |
A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity |
| title_sort |
novel activating effect of the regulatory subunit of protein kinase a on catalytic subunit activity |
| publishDate |
2008 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00039861_v480_n2_p95_Rinaldi http://hdl.handle.net/20.500.12110/paper_00039861_v480_n2_p95_Rinaldi |
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