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...
Guardado en:
| Autores principales: | , , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00039861_v480_n2_p95_Rinaldi |
| Aporte de: |
| id |
todo:paper_00039861_v480_n2_p95_Rinaldi |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_00039861_v480_n2_p95_Rinaldi2023-10-03T13:57:03Z A novel activating effect of the regulatory subunit of protein kinase A on catalytic subunit activity Rinaldi, J. Ocampo, J. Rossi, S. Moreno, S. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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, J. Ocampo, J. Rossi, S. Moreno, S. 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. |
| format |
JOUR |
| author |
Rinaldi, J. Ocampo, J. Rossi, S. Moreno, S. |
| author_facet |
Rinaldi, J. Ocampo, J. Rossi, S. Moreno, S. |
| author_sort |
Rinaldi, J. |
| 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 |
| url |
http://hdl.handle.net/20.500.12110/paper_00039861_v480_n2_p95_Rinaldi |
| work_keys_str_mv |
AT rinaldij anovelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT ocampoj anovelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT rossis anovelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT morenos anovelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT rinaldij novelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT ocampoj novelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT rossis novelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity AT morenos novelactivatingeffectoftheregulatorysubunitofproteinkinaseaoncatalyticsubunitactivity |
| _version_ |
1807315786544447488 |