Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C
Despite a generic, highly conserved motor domain, ATP turnover kinetics and their activation by F-actin vary greatly between myosin-2 isoforms. Here, we present a 2.25 Å pre-powerstroke state (ADP⋅VO4) crystal structure of the human nonmuscle myosin-2C motor domain, one of the slowest myosins charac...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2017-12-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/32742 |
| _version_ | 1828165552000991232 |
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| author | Krishna Chinthalapudi Sarah M Heissler Matthias Preller James R Sellers Dietmar J Manstein |
| author_facet | Krishna Chinthalapudi Sarah M Heissler Matthias Preller James R Sellers Dietmar J Manstein |
| author_sort | Krishna Chinthalapudi |
| collection | DOAJ |
| description | Despite a generic, highly conserved motor domain, ATP turnover kinetics and their activation by F-actin vary greatly between myosin-2 isoforms. Here, we present a 2.25 Å pre-powerstroke state (ADP⋅VO4) crystal structure of the human nonmuscle myosin-2C motor domain, one of the slowest myosins characterized. In combination with integrated mutagenesis, ensemble-solution kinetics, and molecular dynamics simulation approaches, the structure reveals an allosteric communication pathway that connects the distal end of the motor domain with the active site. Disruption of this pathway by mutation of hub residue R788, which forms the center of a cluster of interactions connecting the converter, the SH1-SH2 helix, the relay helix, and the lever, abolishes nonmuscle myosin-2 specific kinetic signatures. Our results provide insights into structural changes in the myosin motor domain that are triggered upon F-actin binding and contribute critically to the mechanochemical behavior of stress fibers, actin arcs, and cortical actin-based structures. |
| first_indexed | 2024-04-12T01:46:36Z |
| format | Article |
| id | doaj.art-e86762993a7243febaacd1f1f457d1c2 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T01:46:36Z |
| publishDate | 2017-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-e86762993a7243febaacd1f1f457d1c22022-12-22T03:53:04ZengeLife Sciences Publications LtdeLife2050-084X2017-12-01610.7554/eLife.32742Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2CKrishna Chinthalapudi0Sarah M Heissler1Matthias Preller2https://orcid.org/0000-0002-7784-4012James R Sellers3https://orcid.org/0000-0001-6296-564XDietmar J Manstein4https://orcid.org/0000-0003-0763-0147Institute for Biophysical Chemistry, OE4350, Hannover Medical School, Hannover, Germany; Division for Structural Biochemistry, OE8830, Hannover Medical School, Hannover, Germany; Cell Adhesion Laboratory, Department of Integrative Structural and Computational Biology, The Scripps Research Institute, Jupiter, United StatesInstitute for Biophysical Chemistry, OE4350, Hannover Medical School, Hannover, Germany; Laboratory of Molecular Physiology, NHLBI, National Institutes of Health, Bethesda, United StatesInstitute for Biophysical Chemistry, OE4350, Hannover Medical School, Hannover, Germany; Centre for Structural Systems Biology (CSSB), German Electron Synchrotron (DESY), Hamburg, GermanyLaboratory of Molecular Physiology, NHLBI, National Institutes of Health, Bethesda, United StatesInstitute for Biophysical Chemistry, OE4350, Hannover Medical School, Hannover, Germany; Division for Structural Biochemistry, OE8830, Hannover Medical School, Hannover, GermanyDespite a generic, highly conserved motor domain, ATP turnover kinetics and their activation by F-actin vary greatly between myosin-2 isoforms. Here, we present a 2.25 Å pre-powerstroke state (ADP⋅VO4) crystal structure of the human nonmuscle myosin-2C motor domain, one of the slowest myosins characterized. In combination with integrated mutagenesis, ensemble-solution kinetics, and molecular dynamics simulation approaches, the structure reveals an allosteric communication pathway that connects the distal end of the motor domain with the active site. Disruption of this pathway by mutation of hub residue R788, which forms the center of a cluster of interactions connecting the converter, the SH1-SH2 helix, the relay helix, and the lever, abolishes nonmuscle myosin-2 specific kinetic signatures. Our results provide insights into structural changes in the myosin motor domain that are triggered upon F-actin binding and contribute critically to the mechanochemical behavior of stress fibers, actin arcs, and cortical actin-based structures.https://elifesciences.org/articles/32742myosincytoskeletonactinallosterySpodoptera frugiperdaArgopecten irradians |
| spellingShingle | Krishna Chinthalapudi Sarah M Heissler Matthias Preller James R Sellers Dietmar J Manstein Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C eLife myosin cytoskeleton actin allostery Spodoptera frugiperda Argopecten irradians |
| title | Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C |
| title_full | Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C |
| title_fullStr | Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C |
| title_full_unstemmed | Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C |
| title_short | Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C |
| title_sort | mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin 2c |
| topic | myosin cytoskeleton actin allostery Spodoptera frugiperda Argopecten irradians |
| url | https://elifesciences.org/articles/32742 |
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