Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin
The auto-inhibited, super-relaxed (SRX) state of cardiac myosin is thought to be crucial for regulating contraction, relaxation, and energy conservation in the heart. We used single ATP turnover experiments to demonstrate that a dilated cardiomyopathy (DCM) mutation (E525K) in human beta-cardiac myo...
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| Format: | Article |
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eLife Sciences Publications Ltd
2022-11-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/77415 |
| _version_ | 1811189896506769408 |
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| author | David V Rasicci Prince Tiwari Skylar ML Bodt Rohini Desetty Fredrik R Sadler Sivaraj Sivaramakrishnan Roger Craig Christopher M Yengo |
| author_facet | David V Rasicci Prince Tiwari Skylar ML Bodt Rohini Desetty Fredrik R Sadler Sivaraj Sivaramakrishnan Roger Craig Christopher M Yengo |
| author_sort | David V Rasicci |
| collection | DOAJ |
| description | The auto-inhibited, super-relaxed (SRX) state of cardiac myosin is thought to be crucial for regulating contraction, relaxation, and energy conservation in the heart. We used single ATP turnover experiments to demonstrate that a dilated cardiomyopathy (DCM) mutation (E525K) in human beta-cardiac myosin increases the fraction of myosin heads in the SRX state (with slow ATP turnover), especially in physiological ionic strength conditions. We also utilized FRET between a C-terminal GFP tag on the myosin tail and Cy3ATP bound to the active site of the motor domain to estimate the fraction of heads in the closed, interacting-heads motif (IHM); we found a strong correlation between the IHM and SRX state. Negative stain electron microscopy and 2D class averaging of the construct demonstrated that the E525K mutation increased the fraction of molecules adopting the IHM. Overall, our results demonstrate that the E525K DCM mutation may reduce muscle force and power by stabilizing the auto-inhibited SRX state. Our studies also provide direct evidence for a correlation between the SRX biochemical state and the IHM structural state in cardiac muscle myosin. Furthermore, the E525 residue may be implicated in crucial electrostatic interactions that modulate this conserved, auto-inhibited conformation of myosin. |
| first_indexed | 2024-04-11T14:42:32Z |
| format | Article |
| id | doaj.art-2a972567fd55436c8074580ba6a5c3a8 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T14:42:32Z |
| publishDate | 2022-11-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
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| spelling | doaj.art-2a972567fd55436c8074580ba6a5c3a82022-12-22T04:17:48ZengeLife Sciences Publications LtdeLife2050-084X2022-11-011110.7554/eLife.77415Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosinDavid V Rasicci0Prince Tiwari1Skylar ML Bodt2Rohini Desetty3Fredrik R Sadler4Sivaraj Sivaramakrishnan5https://orcid.org/0000-0002-9541-6994Roger Craig6https://orcid.org/0000-0002-9707-5409Christopher M Yengo7https://orcid.org/0000-0003-3987-9019Department of Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, United StatesDepartment of Radiology, Division of Cell Biology and Imaging, UMass Chan Medical School, Worcester, United StatesDepartment of Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, United StatesDepartment of Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, United StatesDepartment of Genetics, Cell Biology, and Development, University of Minnesota Twin Cities, Minneapolis, United StatesDepartment of Genetics, Cell Biology, and Development, University of Minnesota Twin Cities, Minneapolis, United StatesDepartment of Radiology, Division of Cell Biology and Imaging, UMass Chan Medical School, Worcester, United StatesDepartment of Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, United StatesThe auto-inhibited, super-relaxed (SRX) state of cardiac myosin is thought to be crucial for regulating contraction, relaxation, and energy conservation in the heart. We used single ATP turnover experiments to demonstrate that a dilated cardiomyopathy (DCM) mutation (E525K) in human beta-cardiac myosin increases the fraction of myosin heads in the SRX state (with slow ATP turnover), especially in physiological ionic strength conditions. We also utilized FRET between a C-terminal GFP tag on the myosin tail and Cy3ATP bound to the active site of the motor domain to estimate the fraction of heads in the closed, interacting-heads motif (IHM); we found a strong correlation between the IHM and SRX state. Negative stain electron microscopy and 2D class averaging of the construct demonstrated that the E525K mutation increased the fraction of molecules adopting the IHM. Overall, our results demonstrate that the E525K DCM mutation may reduce muscle force and power by stabilizing the auto-inhibited SRX state. Our studies also provide direct evidence for a correlation between the SRX biochemical state and the IHM structural state in cardiac muscle myosin. Furthermore, the E525 residue may be implicated in crucial electrostatic interactions that modulate this conserved, auto-inhibited conformation of myosin.https://elifesciences.org/articles/77415cardiac muscle myosinFRETsuper-relaxed stateinteracting-heads motifdilated cardiomyopathy |
| spellingShingle | David V Rasicci Prince Tiwari Skylar ML Bodt Rohini Desetty Fredrik R Sadler Sivaraj Sivaramakrishnan Roger Craig Christopher M Yengo Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin eLife cardiac muscle myosin FRET super-relaxed state interacting-heads motif dilated cardiomyopathy |
| title | Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin |
| title_full | Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin |
| title_fullStr | Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin |
| title_full_unstemmed | Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin |
| title_short | Dilated cardiomyopathy mutation E525K in human beta-cardiac myosin stabilizes the interacting-heads motif and super-relaxed state of myosin |
| title_sort | dilated cardiomyopathy mutation e525k in human beta cardiac myosin stabilizes the interacting heads motif and super relaxed state of myosin |
| topic | cardiac muscle myosin FRET super-relaxed state interacting-heads motif dilated cardiomyopathy |
| url | https://elifesciences.org/articles/77415 |
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