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Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome
Overproduction of cortisol by the hypothalamus–pituitary–adrenal hormone system results in the clinical disorder known as Cushing's syndrome. Genomics studies have identified a key mutation (L205R) in the α‐isoform of the catalytic subunit of cAMP‐dependent protein kinase (PKACα) in adrenal ade...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881547/ https://www.ncbi.nlm.nih.gov/pubmed/29632813 http://dx.doi.org/10.1002/2211-5463.12396 |
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author | Luzi, Nicole M. Lyons, Charles E. Peterson, Darrell L. Ellis, Keith C. |
author_facet | Luzi, Nicole M. Lyons, Charles E. Peterson, Darrell L. Ellis, Keith C. |
author_sort | Luzi, Nicole M. |
collection | PubMed |
description | Overproduction of cortisol by the hypothalamus–pituitary–adrenal hormone system results in the clinical disorder known as Cushing's syndrome. Genomics studies have identified a key mutation (L205R) in the α‐isoform of the catalytic subunit of cAMP‐dependent protein kinase (PKACα) in adrenal adenomas of patients with adrenocorticotropic hormone‐independent Cushing's syndrome. Here, we conducted kinetics and inhibition studies on the L205R‐PKACα mutant. We have found that the L205R mutation affects the kinetics of both Kemptide and ATP as substrates, decreasing the catalytic efficiency (k (cat)/K (M)) for each substrate by 12‐fold and 4.5‐fold, respectively. We have also determined the IC (50) and K (i) for the peptide substrate‐competitive inhibitor PKI(5–24) and the ATP‐competitive inhibitor H89. The L205R mutation had no effect on the potency of H89, but causes a > 250‐fold loss in potency for PKI(5–24). Collectively, these data provide insights for the development of L205R‐PKACα inhibitors as potential therapeutics. |
format | Online Article Text |
id | pubmed-5881547 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58815472018-04-09 Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome Luzi, Nicole M. Lyons, Charles E. Peterson, Darrell L. Ellis, Keith C. FEBS Open Bio Research Articles Overproduction of cortisol by the hypothalamus–pituitary–adrenal hormone system results in the clinical disorder known as Cushing's syndrome. Genomics studies have identified a key mutation (L205R) in the α‐isoform of the catalytic subunit of cAMP‐dependent protein kinase (PKACα) in adrenal adenomas of patients with adrenocorticotropic hormone‐independent Cushing's syndrome. Here, we conducted kinetics and inhibition studies on the L205R‐PKACα mutant. We have found that the L205R mutation affects the kinetics of both Kemptide and ATP as substrates, decreasing the catalytic efficiency (k (cat)/K (M)) for each substrate by 12‐fold and 4.5‐fold, respectively. We have also determined the IC (50) and K (i) for the peptide substrate‐competitive inhibitor PKI(5–24) and the ATP‐competitive inhibitor H89. The L205R mutation had no effect on the potency of H89, but causes a > 250‐fold loss in potency for PKI(5–24). Collectively, these data provide insights for the development of L205R‐PKACα inhibitors as potential therapeutics. John Wiley and Sons Inc. 2018-03-11 /pmc/articles/PMC5881547/ /pubmed/29632813 http://dx.doi.org/10.1002/2211-5463.12396 Text en © 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Articles Luzi, Nicole M. Lyons, Charles E. Peterson, Darrell L. Ellis, Keith C. Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title | Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title_full | Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title_fullStr | Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title_full_unstemmed | Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title_short | Kinetics and inhibition studies of the L205R mutant of cAMP‐dependent protein kinase involved in Cushing's syndrome |
title_sort | kinetics and inhibition studies of the l205r mutant of camp‐dependent protein kinase involved in cushing's syndrome |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5881547/ https://www.ncbi.nlm.nih.gov/pubmed/29632813 http://dx.doi.org/10.1002/2211-5463.12396 |
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