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Insights into the Phosphoryl Transfer Catalyzed by cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study of Complexes with Various Metals and Peptide Substrate SP20
[Image: see text] X-ray structures of several ternary substrate and product complexes of the catalytic subunit of cAMP-dependent protein kinase (PKAc) have been determined with different bound metal ions. In the PKAc complexes, Mg(2+), Ca(2+), Sr(2+), and Ba(2+) metal ions could bind to the active s...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2013
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3666212/ https://www.ncbi.nlm.nih.gov/pubmed/23672593 http://dx.doi.org/10.1021/bi400066a |
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author | Gerlits, Oksana Waltman, Mary Jo Taylor, Susan Langan, Paul Kovalevsky, Andrey |
author_facet | Gerlits, Oksana Waltman, Mary Jo Taylor, Susan Langan, Paul Kovalevsky, Andrey |
author_sort | Gerlits, Oksana |
collection | PubMed |
description | [Image: see text] X-ray structures of several ternary substrate and product complexes of the catalytic subunit of cAMP-dependent protein kinase (PKAc) have been determined with different bound metal ions. In the PKAc complexes, Mg(2+), Ca(2+), Sr(2+), and Ba(2+) metal ions could bind to the active site and facilitate the phosphoryl transfer reaction. ATP and a substrate peptide (SP20) were modified, and the reaction products ADP and the phosphorylated peptide were found trapped in the enzyme active site. Finally, we determined the structure of a pseudo-Michaelis complex containing Mg(2+), nonhydrolyzable AMP-PCP (β,γ-methyleneadenosine 5′-triphosphate) and SP20. The product structures together with the pseudo-Michaelis complex provide snapshots of different stages of the phosphorylation reaction. Comparison of these structures reveals conformational, coordination, and hydrogen bonding changes that might occur during the reaction and shed new light on its mechanism, roles of metals, and active site residues. |
format | Online Article Text |
id | pubmed-3666212 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-36662122013-05-29 Insights into the Phosphoryl Transfer Catalyzed by cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study of Complexes with Various Metals and Peptide Substrate SP20 Gerlits, Oksana Waltman, Mary Jo Taylor, Susan Langan, Paul Kovalevsky, Andrey Biochemistry [Image: see text] X-ray structures of several ternary substrate and product complexes of the catalytic subunit of cAMP-dependent protein kinase (PKAc) have been determined with different bound metal ions. In the PKAc complexes, Mg(2+), Ca(2+), Sr(2+), and Ba(2+) metal ions could bind to the active site and facilitate the phosphoryl transfer reaction. ATP and a substrate peptide (SP20) were modified, and the reaction products ADP and the phosphorylated peptide were found trapped in the enzyme active site. Finally, we determined the structure of a pseudo-Michaelis complex containing Mg(2+), nonhydrolyzable AMP-PCP (β,γ-methyleneadenosine 5′-triphosphate) and SP20. The product structures together with the pseudo-Michaelis complex provide snapshots of different stages of the phosphorylation reaction. Comparison of these structures reveals conformational, coordination, and hydrogen bonding changes that might occur during the reaction and shed new light on its mechanism, roles of metals, and active site residues. American Chemical Society 2013-05-01 2013-05-28 /pmc/articles/PMC3666212/ /pubmed/23672593 http://dx.doi.org/10.1021/bi400066a Text en Copyright © 2013 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) |
spellingShingle | Gerlits, Oksana Waltman, Mary Jo Taylor, Susan Langan, Paul Kovalevsky, Andrey Insights into the Phosphoryl Transfer Catalyzed by cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study of Complexes with Various Metals and Peptide Substrate SP20 |
title | Insights into the Phosphoryl Transfer Catalyzed by
cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study
of Complexes with Various Metals and Peptide Substrate SP20 |
title_full | Insights into the Phosphoryl Transfer Catalyzed by
cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study
of Complexes with Various Metals and Peptide Substrate SP20 |
title_fullStr | Insights into the Phosphoryl Transfer Catalyzed by
cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study
of Complexes with Various Metals and Peptide Substrate SP20 |
title_full_unstemmed | Insights into the Phosphoryl Transfer Catalyzed by
cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study
of Complexes with Various Metals and Peptide Substrate SP20 |
title_short | Insights into the Phosphoryl Transfer Catalyzed by
cAMP-Dependent Protein Kinase: An X-ray Crystallographic Study
of Complexes with Various Metals and Peptide Substrate SP20 |
title_sort | insights into the phosphoryl transfer catalyzed by
camp-dependent protein kinase: an x-ray crystallographic study
of complexes with various metals and peptide substrate sp20 |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3666212/ https://www.ncbi.nlm.nih.gov/pubmed/23672593 http://dx.doi.org/10.1021/bi400066a |
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