Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability
CaMKIIα plays a fundamental role in learning and memory and is a key determinant of synaptic plasticity. Its kinase activity is regulated by the binding of Ca2+/CaM and by autophosphorylation that operates in an activity-dependent manner. Though many mutations in CAMK2A were linked to a variety of n...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2022.970031/full |
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| author | Hajime Fujii Hiroyuki Kidokoro Yayoi Kondo Masahiro Kawaguchi Shin-ichiro Horigane Shin-ichiro Horigane Jun Natsume Jun Natsume Sayaka Takemoto-Kimura Sayaka Takemoto-Kimura Haruhiko Bito |
| author_facet | Hajime Fujii Hiroyuki Kidokoro Yayoi Kondo Masahiro Kawaguchi Shin-ichiro Horigane Shin-ichiro Horigane Jun Natsume Jun Natsume Sayaka Takemoto-Kimura Sayaka Takemoto-Kimura Haruhiko Bito |
| author_sort | Hajime Fujii |
| collection | DOAJ |
| description | CaMKIIα plays a fundamental role in learning and memory and is a key determinant of synaptic plasticity. Its kinase activity is regulated by the binding of Ca2+/CaM and by autophosphorylation that operates in an activity-dependent manner. Though many mutations in CAMK2A were linked to a variety of neurological disorders, the multiplicity of its functional substrates renders the systematic molecular phenotyping challenging. In this study, we report a new case of CAMK2A P212L, a recurrent mutation, in a patient with an intellectual disability. To quantify the effect of this mutation, we developed a FRET-based kinase phenotyping strategy and measured aberrance in Ca2+/CaM-dependent activation dynamics in vitro and in synaptically connected neurons. CaMKIIα P212L revealed a significantly facilitated Ca2+/CaM-dependent activation in vitro. Consistently, this mutant showed faster activation and more delayed inactivation in neurons. More prolonged kinase activation was also accompanied by a leftward shift in the CaMKIIα input frequency tuning curve. In keeping with this, molecular phenotyping of other reported CAMK2A de novo mutations linked to intellectual disability revealed aberrant facilitation of Ca2+/CaM-dependent activation of CaMKIIα in most cases. Finally, the pharmacological reversal of CAMK2A P212L phenotype in neurons was demonstrated using an FDA-approved NMDA receptor antagonist memantine, providing a basis for targeted therapeutics in CAMK2A-linked intellectual disability. Taken together, FRET-based kinase mutation phenotyping sheds light on the biological impact of CAMK2A mutations and provides a selective, sensitive, quantitative, and scalable strategy for gaining novel insights into the molecular etiology of intellectual disability. |
| first_indexed | 2024-04-11T14:13:46Z |
| format | Article |
| id | doaj.art-a47257885bb04a219a003ae87b8317e3 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-04-11T14:13:46Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-a47257885bb04a219a003ae87b8317e32022-12-22T04:19:36ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992022-09-011510.3389/fnmol.2022.970031970031Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disabilityHajime Fujii0Hiroyuki Kidokoro1Yayoi Kondo2Masahiro Kawaguchi3Shin-ichiro Horigane4Shin-ichiro Horigane5Jun Natsume6Jun Natsume7Sayaka Takemoto-Kimura8Sayaka Takemoto-Kimura9Haruhiko Bito10Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, JapanDepartment of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, JapanDepartment of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Neuroscience I, Research Institute of Environmental Medicine (RIEM), Nagoya University, Nagoya, JapanDepartment of Molecular/Cellular Neuroscience, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Developmental Disability Medicine, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Neuroscience I, Research Institute of Environmental Medicine (RIEM), Nagoya University, Nagoya, JapanDepartment of Molecular/Cellular Neuroscience, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Tokyo, JapanCaMKIIα plays a fundamental role in learning and memory and is a key determinant of synaptic plasticity. Its kinase activity is regulated by the binding of Ca2+/CaM and by autophosphorylation that operates in an activity-dependent manner. Though many mutations in CAMK2A were linked to a variety of neurological disorders, the multiplicity of its functional substrates renders the systematic molecular phenotyping challenging. In this study, we report a new case of CAMK2A P212L, a recurrent mutation, in a patient with an intellectual disability. To quantify the effect of this mutation, we developed a FRET-based kinase phenotyping strategy and measured aberrance in Ca2+/CaM-dependent activation dynamics in vitro and in synaptically connected neurons. CaMKIIα P212L revealed a significantly facilitated Ca2+/CaM-dependent activation in vitro. Consistently, this mutant showed faster activation and more delayed inactivation in neurons. More prolonged kinase activation was also accompanied by a leftward shift in the CaMKIIα input frequency tuning curve. In keeping with this, molecular phenotyping of other reported CAMK2A de novo mutations linked to intellectual disability revealed aberrant facilitation of Ca2+/CaM-dependent activation of CaMKIIα in most cases. Finally, the pharmacological reversal of CAMK2A P212L phenotype in neurons was demonstrated using an FDA-approved NMDA receptor antagonist memantine, providing a basis for targeted therapeutics in CAMK2A-linked intellectual disability. Taken together, FRET-based kinase mutation phenotyping sheds light on the biological impact of CAMK2A mutations and provides a selective, sensitive, quantitative, and scalable strategy for gaining novel insights into the molecular etiology of intellectual disability.https://www.frontiersin.org/articles/10.3389/fnmol.2022.970031/fullCaMKIIintellectual disabilityneurodevelopmental disordersimagingFRETde novo mutation |
| spellingShingle | Hajime Fujii Hiroyuki Kidokoro Yayoi Kondo Masahiro Kawaguchi Shin-ichiro Horigane Shin-ichiro Horigane Jun Natsume Jun Natsume Sayaka Takemoto-Kimura Sayaka Takemoto-Kimura Haruhiko Bito Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability Frontiers in Molecular Neuroscience CaMKII intellectual disability neurodevelopmental disorders imaging FRET de novo mutation |
| title | Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability |
| title_full | Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability |
| title_fullStr | Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability |
| title_full_unstemmed | Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability |
| title_short | Förster resonance energy transfer-based kinase mutation phenotyping reveals an aberrant facilitation of Ca2+/calmodulin-dependent CaMKIIα activity in de novo mutations related to intellectual disability |
| title_sort | forster resonance energy transfer based kinase mutation phenotyping reveals an aberrant facilitation of ca2 calmodulin dependent camkiiα activity in de novo mutations related to intellectual disability |
| topic | CaMKII intellectual disability neurodevelopmental disorders imaging FRET de novo mutation |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2022.970031/full |
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