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GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain
Ca(2+)/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) is a key neuronal signaling protein and an emerging drug target. The central hub domain regulates the activity of CaMKIIα by organizing the holoenzyme complex into functional oligomers, yet pharmacological modulation of the hub do...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8346900/ https://www.ncbi.nlm.nih.gov/pubmed/34330837 http://dx.doi.org/10.1073/pnas.2108079118 |
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author | Leurs, Ulrike Klein, Anders B. McSpadden, Ethan D. Griem-Krey, Nane Solbak, Sara M. Ø. Houlton, Josh Villumsen, Inge S. Vogensen, Stine B. Hamborg, Louise Gauger, Stine J. Palmelund, Line B. Larsen, Anne Sofie G. Shehata, Mohamed A. Kelstrup, Christian D. Olsen, Jesper V. Bach, Anders Burnie, Robert O. Kerr, D. Steven Gowing, Emma K. Teurlings, Selina M. W. Chi, Chris C. Gee, Christine L. Frølund, Bente Kornum, Birgitte R. van Woerden, Geeske M. Clausen, Rasmus P. Kuriyan, John Clarkson, Andrew N. Wellendorph, Petrine |
author_facet | Leurs, Ulrike Klein, Anders B. McSpadden, Ethan D. Griem-Krey, Nane Solbak, Sara M. Ø. Houlton, Josh Villumsen, Inge S. Vogensen, Stine B. Hamborg, Louise Gauger, Stine J. Palmelund, Line B. Larsen, Anne Sofie G. Shehata, Mohamed A. Kelstrup, Christian D. Olsen, Jesper V. Bach, Anders Burnie, Robert O. Kerr, D. Steven Gowing, Emma K. Teurlings, Selina M. W. Chi, Chris C. Gee, Christine L. Frølund, Bente Kornum, Birgitte R. van Woerden, Geeske M. Clausen, Rasmus P. Kuriyan, John Clarkson, Andrew N. Wellendorph, Petrine |
author_sort | Leurs, Ulrike |
collection | PubMed |
description | Ca(2+)/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) is a key neuronal signaling protein and an emerging drug target. The central hub domain regulates the activity of CaMKIIα by organizing the holoenzyme complex into functional oligomers, yet pharmacological modulation of the hub domain has never been demonstrated. Here, using a combination of photoaffinity labeling and chemical proteomics, we show that compounds related to the natural substance γ-hydroxybutyrate (GHB) bind selectively to CaMKIIα. By means of a 2.2-Å x-ray crystal structure of ligand-bound CaMKIIα hub, we reveal the molecular details of the binding site deep within the hub. Furthermore, we show that binding of GHB and related analogs to this site promotes concentration-dependent increases in hub thermal stability believed to alter holoenzyme functionality. Selectively under states of pathological CaMKIIα activation, hub ligands provide a significant and sustained neuroprotection, which is both time and dose dependent. This is demonstrated in neurons exposed to excitotoxicity and in a mouse model of cerebral ischemia with the selective GHB analog, HOCPCA (3-hydroxycyclopent-1-enecarboxylic acid). Together, our results indicate a hitherto unknown mechanism for neuroprotection by a highly specific and unforeseen interaction between the CaMKIIα hub domain and small molecule brain-penetrant GHB analogs. This establishes GHB analogs as powerful tools for investigating CaMKII neuropharmacology in general and as potential therapeutic compounds for cerebral ischemia in particular. |
format | Online Article Text |
id | pubmed-8346900 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-83469002021-08-23 GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain Leurs, Ulrike Klein, Anders B. McSpadden, Ethan D. Griem-Krey, Nane Solbak, Sara M. Ø. Houlton, Josh Villumsen, Inge S. Vogensen, Stine B. Hamborg, Louise Gauger, Stine J. Palmelund, Line B. Larsen, Anne Sofie G. Shehata, Mohamed A. Kelstrup, Christian D. Olsen, Jesper V. Bach, Anders Burnie, Robert O. Kerr, D. Steven Gowing, Emma K. Teurlings, Selina M. W. Chi, Chris C. Gee, Christine L. Frølund, Bente Kornum, Birgitte R. van Woerden, Geeske M. Clausen, Rasmus P. Kuriyan, John Clarkson, Andrew N. Wellendorph, Petrine Proc Natl Acad Sci U S A Biological Sciences Ca(2+)/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) is a key neuronal signaling protein and an emerging drug target. The central hub domain regulates the activity of CaMKIIα by organizing the holoenzyme complex into functional oligomers, yet pharmacological modulation of the hub domain has never been demonstrated. Here, using a combination of photoaffinity labeling and chemical proteomics, we show that compounds related to the natural substance γ-hydroxybutyrate (GHB) bind selectively to CaMKIIα. By means of a 2.2-Å x-ray crystal structure of ligand-bound CaMKIIα hub, we reveal the molecular details of the binding site deep within the hub. Furthermore, we show that binding of GHB and related analogs to this site promotes concentration-dependent increases in hub thermal stability believed to alter holoenzyme functionality. Selectively under states of pathological CaMKIIα activation, hub ligands provide a significant and sustained neuroprotection, which is both time and dose dependent. This is demonstrated in neurons exposed to excitotoxicity and in a mouse model of cerebral ischemia with the selective GHB analog, HOCPCA (3-hydroxycyclopent-1-enecarboxylic acid). Together, our results indicate a hitherto unknown mechanism for neuroprotection by a highly specific and unforeseen interaction between the CaMKIIα hub domain and small molecule brain-penetrant GHB analogs. This establishes GHB analogs as powerful tools for investigating CaMKII neuropharmacology in general and as potential therapeutic compounds for cerebral ischemia in particular. National Academy of Sciences 2021-08-03 2021-07-30 /pmc/articles/PMC8346900/ /pubmed/34330837 http://dx.doi.org/10.1073/pnas.2108079118 Text en Copyright © 2021 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences Leurs, Ulrike Klein, Anders B. McSpadden, Ethan D. Griem-Krey, Nane Solbak, Sara M. Ø. Houlton, Josh Villumsen, Inge S. Vogensen, Stine B. Hamborg, Louise Gauger, Stine J. Palmelund, Line B. Larsen, Anne Sofie G. Shehata, Mohamed A. Kelstrup, Christian D. Olsen, Jesper V. Bach, Anders Burnie, Robert O. Kerr, D. Steven Gowing, Emma K. Teurlings, Selina M. W. Chi, Chris C. Gee, Christine L. Frølund, Bente Kornum, Birgitte R. van Woerden, Geeske M. Clausen, Rasmus P. Kuriyan, John Clarkson, Andrew N. Wellendorph, Petrine GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title | GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title_full | GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title_fullStr | GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title_full_unstemmed | GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title_short | GHB analogs confer neuroprotection through specific interaction with the CaMKIIα hub domain |
title_sort | ghb analogs confer neuroprotection through specific interaction with the camkiiα hub domain |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8346900/ https://www.ncbi.nlm.nih.gov/pubmed/34330837 http://dx.doi.org/10.1073/pnas.2108079118 |
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