Deciphering the super relaxed state of human β-cardiac myosin and the mode of action of mavacamten from myosin molecules to muscle fibers

Anderson, Robert L.; Trivedi, Darshan V.; Sarkar, Saswata S.; Henze, Marcus; Ma, Weikang; Gong, Henry; Rogers, Christopher S.; Gorham, Joshua M.; Wong, Fiona L.; Morck, Makenna M.; Seidman, Jonathan G.; Ruppel, Kathleen M.; Irving, Thomas C.; Cooke, Roger; Green, Eric M.; Spudich, James A.

doi:10.1073/pnas.1809540115

Title: Deciphering the super relaxed state of human β-cardiac myosin and the mode of action of mavacamten from myosin molecules to muscle fibers

Journal Article · Mon Aug 13 00:00:00 EDT 2018 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1809540115· OSTI ID:1464331

Anderson, Robert L.; Trivedi, Darshan V.; Sarkar, Saswata S.; Henze, Marcus; Ma, Weikang; Gong, Henry; Rogers, Christopher S.; Gorham, Joshua M.; Wong, Fiona L.; Morck, Makenna M.; Seidman, Jonathan G.; Ruppel, Kathleen M.; Irving, Thomas C.; Cooke, Roger; Green, Eric M.; Spudich, James A.

Mutations in β-cardiac myosin, the predominant motor protein for human heart contraction, can alter power output and cause cardiomyopathy. However, measurements of the intrinsic force, velocity, and ATPase activity of myosin have not provided a consistent mechanism to link mutations to muscle pathology. An alternative model posits that mutations in myosin affect the stability of a sequestered, super relaxed state (SRX) of the protein with very slow ATP hydrolysis and thereby change the number of myosin heads accessible to actin. Here we show that purified human β-cardiac myosin exists partly in an SRX and may in part correspond to a folded-back conformation of myosin heads observed in muscle fibers around the thick filament backbone. Mutations that cause hypertrophic cardiomyopathy destabilize this state, while the small molecule mavacamten promotes it. Furthermore, these findings provide a biochemical and structural link between the genetics and physiology of cardiomyopathy with implications for therapeutic strategies.

View Journal Article

Cite

Export

Save

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-06CH11357; GM33289; HL117138; AR062279; HL084553; P41 GM103622

OSTI ID:: 1464331

Alternate ID(s):: OSTI ID: 1470476

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Vol. 115 Journal Issue: 35; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 175 works

Citation information provided by
Web of Science

References (64)

Blebbistatin Stabilizes the Helical Order of Myosin Filaments by Promoting the Switch 2 Closed State Zhao, Fa-Qing; Padrón, Raúl; Craig, Roger Biophysical Journal, Vol. 95, Issue 7 https://doi.org/10.1529/biophysj.108.137067	journal	October 2008
Atomic model of a myosin filament in the relaxed state Woodhead, John L.; Zhao, Fa-Qing; Craig, Roger Nature, Vol. 436, Issue 7054 https://doi.org/10.1038/nature03920	journal	August 2005
Helical Order in Tarantula Thick Filaments Requires the “Closed” Conformation of the Myosin Head Zoghbi, M. E.; Woodhead, J. L.; Craig, R. Journal of Molecular Biology, Vol. 342, Issue 4 https://doi.org/10.1016/j.jmb.2004.07.037	journal	September 2004
The myosin mesa and a possible unifying hypothesis for the molecular basis of human hypertrophic cardiomyopathy Spudich, James A. Biochemical Society Transactions, Vol. 43, Issue 1 https://doi.org/10.1042/BST20140324	journal	January 2015
Myosin-binding protein C corrects an intrinsic inhomogeneity in cardiac excitation-contraction coupling Previs, Michael J.; Prosser, Benjamin L.; Mun, Ji Young Science Advances, Vol. 1, Issue 1 https://doi.org/10.1126/sciadv.1400205	journal	February 2015
Light-chain phosphorylation controls the conformation of vertebrate non-muscle and smooth muscle myosin molecules Craig, Roger; Smith, Robin; Kendrick-Jones, John Nature, Vol. 302, Issue 5907 https://doi.org/10.1038/302436a0	journal	March 1983
Time-resolved X-ray diffraction studies of the myosin layer-line reflections during muscle contraction Huxley, H. E.; Faruqi, A. R.; Kress, M. Journal of Molecular Biology, Vol. 158, Issue 4 https://doi.org/10.1016/0022-2836(82)90253-4	journal	July 1982
Spectroscopic Studies of the Super Relaxed State of Skeletal Muscle Nogara, Leonardo; Naber, Nariman; Pate, Edward PLOS ONE, Vol. 11, Issue 8 https://doi.org/10.1371/journal.pone.0160100	journal	August 2016
Calcium-dependent structural changes in scallop heavy meromyosin Stafford, W. F.; Jacobsen, M. P.; Woodhead, J. Journal of Molecular Biology, Vol. 307, Issue 1 https://doi.org/10.1006/jmbi.2000.4490	journal	March 2001
The Filament Lattice of Striated Muscle Millman, Barry M. Physiological Reviews, Vol. 78, Issue 2 https://doi.org/10.1152/physrev.1998.78.2.359	journal	April 1998
Slow Myosin ATP Turnover in the Super-Relaxed State in Tarantula Muscle Naber, Nariman; Cooke, Roger; Pate, Edward Journal of Molecular Biology, Vol. 411, Issue 5 https://doi.org/10.1016/j.jmb.2011.06.051	journal	September 2011
Crystal structures of human cardiac beta-myosin II S2- provide insight into the functional role of the S2 subfragment Blankenfeldt, W.; Thoma, N. H.; Wray, J. S. Proceedings of the National Academy of Sciences, Vol. 103, Issue 47 https://doi.org/10.1073/pnas.0606741103	journal	November 2006
Myosin subfragment-1 is sufficient to move actin filaments in vitro Toyoshima, Yoko Yano; Kron, Stephen J.; McNally, Elizabeth M. Nature, Vol. 328, Issue 6130 https://doi.org/10.1038/328536a0	journal	August 1987
Three-Dimensional Reconstruction of Tarantula Myosin Filaments Suggests How Phosphorylation May Regulate Myosin Activity Alamo, Lorenzo; Wriggers, Willy; Pinto, Antonio Journal of Molecular Biology, Vol. 384, Issue 4 https://doi.org/10.1016/j.jmb.2008.10.013	journal	December 2008
Structure of the myosin projections on native thick filaments from vertebrate skeletal muscle Knight, Peter; Trinick, John Journal of Molecular Biology, Vol. 177, Issue 3 https://doi.org/10.1016/0022-2836(84)90295-X	journal	August 1984
Omecamtiv mercabil and blebbistatin modulate cardiac contractility by perturbing the regulatory state of the myosin filament: Myosin filament regulation in heart muscle Kampourakis, Thomas; Zhang, Xuemeng; Sun, Yin-Biao The Journal of Physiology, Vol. 596, Issue 1 https://doi.org/10.1113/JP275050	journal	November 2017
Crystal Structure of a Vertebrate Smooth Muscle Myosin Motor Domain and Its Complex with the Essential Light Chain Dominguez, Roberto; Freyzon, Yelena; Trybus, Kathleen M. Cell, Vol. 94, Issue 5 https://doi.org/10.1016/S0092-8674(00)81598-6	journal	September 1998
Atomic model of the human cardiac muscle myosin filament AL-Khayat, H. A.; Kensler, R. W.; Squire, J. M. Proceedings of the National Academy of Sciences, Vol. 110, Issue 1 https://doi.org/10.1073/pnas.1212708110	journal	December 2012
Multidimensional structure-function relationships in human β-cardiac myosin from population-scale genetic variation Homburger, Julian R.; Green, Eric M.; Caleshu, Colleen Proceedings of the National Academy of Sciences, Vol. 113, Issue 24 https://doi.org/10.1073/pnas.1606950113	journal	May 2016
Early-Onset Hypertrophic Cardiomyopathy Mutations Significantly Increase the Velocity, Force, and Actin-Activated ATPase Activity of Human β-Cardiac Myosin Adhikari, Arjun S.; Kooiker, Kristina B.; Sarkar, Saswata S. Cell Reports, Vol. 17, Issue 11 https://doi.org/10.1016/j.celrep.2016.11.040	journal	December 2016
Molecular mechanisms of cardiomyopathy phenotypes associated with myosin light chain mutations Huang, Wenrui; Szczesna-Cordary, Danuta Journal of Muscle Research and Cell Motility, Vol. 36, Issue 6 https://doi.org/10.1007/s10974-015-9423-3	journal	September 2015
A Mouse Model of Familial Hypertrophic Cardiomyopathy Geisterfer-Lowrance, A. A. T.; Christe, M.; Conner, D. A. Science, Vol. 272, Issue 5262 https://doi.org/10.1126/science.272.5262.731	journal	May 1996
Contractility parameters of human β-cardiac myosin with the hypertrophic cardiomyopathy mutation R403Q show loss of motor function Nag, Suman; Sommese, Ruth F.; Ujfalusi, Zoltan Science Advances, Vol. 1, Issue 9 https://doi.org/10.1126/sciadv.1500511	journal	October 2015
Ablation of cardiac myosin binding protein-C disrupts the super-relaxed state of myosin in murine cardiomyocytes McNamara, James W.; Li, Amy; Smith, Nicola J. Journal of Molecular and Cellular Cardiology, Vol. 94 https://doi.org/10.1016/j.yjmcc.2016.03.009	journal	May 2016
Visualization of Head–Head Interactions in the Inhibited State of Smooth Muscle Myosin Wendt, Thomas; Taylor, Dianne; Messier, Terri The Journal of Cell Biology, Vol. 147, Issue 7 https://doi.org/10.1083/jcb.147.7.1385	journal	December 1999
Dynamics of left ventricular emptying in hypertrophic subaortic stenosis Wilson, W. Stanley; Criley, J. Michael; Ross, Richard S. American Heart Journal, Vol. 73, Issue 1 https://doi.org/10.1016/0002-8703(67)90303-1	journal	January 1967
Understanding Cardiomyopathy Phenotypes Based on the Functional Impact of Mutations in the Myosin Motor Moore, Jeffrey R.; Leinwand, Leslie; Warshaw, David M. Circulation Research, Vol. 111, Issue 3 https://doi.org/10.1161/CIRCRESAHA.110.223842	journal	July 2012
Titin strain contributes to the Frank–Starling law of the heart by structural rearrangements of both thin- and thick-filament proteins Ait-Mou, Younss; Hsu, Karen; Farman, Gerrie P. Proceedings of the National Academy of Sciences, Vol. 113, Issue 8 https://doi.org/10.1073/pnas.1516732113	journal	February 2016
Temperature and Ligand Dependence of Conformation and Helical Order in Myosin Filaments Xu, S.; Offer, G.; Gu, J. Biochemistry, Vol. 42, Issue 2 https://doi.org/10.1021/bi026085t	journal	January 2003
The Myosin Inhibitor Blebbistatin Stabilizes the Super-Relaxed State in Skeletal Muscle Wilson, Clyde; Naber, Nariman; Pate, Edward Biophysical Journal, Vol. 107, Issue 7 https://doi.org/10.1016/j.bpj.2014.07.075	journal	October 2014
Role of the Tail in the Regulated State of Myosin 2 Jung, Hyun Suk; Billington, Neil; Thirumurugan, Kavitha Journal of Molecular Biology, Vol. 408, Issue 5 https://doi.org/10.1016/j.jmb.2011.03.019	journal	May 2011
Phosphorylation of myosin regulatory light chain controls myosin head conformation in cardiac muscle Kampourakis, Thomas; Irving, Malcolm Journal of Molecular and Cellular Cardiology, Vol. 85 https://doi.org/10.1016/j.yjmcc.2015.06.002	journal	August 2015
Structures of Smooth Muscle Myosin and Heavy Meromyosin in the Folded, Shutdown State Burgess, Stan A.; Yu, Shuizi; Walker, Matt L. Journal of Molecular Biology, Vol. 372, Issue 5 https://doi.org/10.1016/j.jmb.2007.07.014	journal	October 2007
A molecular basis for familial hypertrophic cardiomyopathy: A β cardiac myosin heavy chain gene missense mutation Geisterfer-Lowrance, Anja A. T.; Kass, Susan; Tanigawa, Gary Cell, Vol. 62, Issue 5 https://doi.org/10.1016/0092-8674(90)90274-I	journal	September 1990
Myosin binding protein-C activates thin filaments and inhibits thick filaments in heart muscle cells Kampourakis, Thomas; Yan, Ziqian; Gautel, Mathias Proceedings of the National Academy of Sciences, Vol. 111, Issue 52 https://doi.org/10.1073/pnas.1413922112	journal	December 2014
The role of super-relaxed myosin in skeletal and cardiac muscle McNamara, James W.; Li, Amy; dos Remedios, Cristobal G. Biophysical Reviews, Vol. 7, Issue 1 https://doi.org/10.1007/s12551-014-0151-5	journal	December 2014
Force generation by skeletal muscle is controlled by mechanosensing in myosin filaments Linari, Marco; Brunello, Elisabetta; Reconditi, Massimo Nature, Vol. 528, Issue 7581 https://doi.org/10.1038/nature15727	journal	November 2015
A small-molecule modulator of cardiac myosin acts on multiple stages of the myosin chemomechanical cycle Kawas, Raja F.; Anderson, Robert L.; Ingle, Sadie R. Bartholomew Journal of Biological Chemistry, Vol. 292, Issue 40 https://doi.org/10.1074/jbc.M117.776815	journal	August 2017
Spare the rod, spoil the regulation: Necessity for a myosin rod Trybus, K. M.; Freyzon, Y.; Faust, L. Z. Proceedings of the National Academy of Sciences, Vol. 94, Issue 1 https://doi.org/10.1073/pnas.94.1.48	journal	January 1997
The role of the myosin ATPase activity in adaptive thermogenesis by skeletal muscle Cooke, Roger Biophysical Reviews, Vol. 3, Issue 1 https://doi.org/10.1007/s12551-011-0044-9	journal	March 2011
A small-molecule inhibitor of sarcomere contractility suppresses hypertrophic cardiomyopathy in mice Green, E. M.; Wakimoto, H.; Anderson, R. L. Science, Vol. 351, Issue 6273 https://doi.org/10.1126/science.aad3456	journal	February 2016
Mavacamten stabilizes an autoinhibited state of two-headed cardiac myosin Rohde, John A.; Roopnarine, Osha; Thomas, David D. Proceedings of the National Academy of Sciences, Vol. 115, Issue 32 https://doi.org/10.1073/pnas.1720342115	journal	July 2018
Three-dimensional structure of vertebrate cardiac muscle myosin filaments Zoghbi, M. E.; Woodhead, J. L.; Moss, R. L. Proceedings of the National Academy of Sciences, Vol. 105, Issue 7 https://doi.org/10.1073/pnas.0708912105	journal	February 2008
Effects of myosin variants on interacting-heads motif explain distinct hypertrophic and dilated cardiomyopathy phenotypes Alamo, Lorenzo; Ware, James S.; Pinto, Antonio eLife, Vol. 6 https://doi.org/10.7554/eLife.24634	journal	June 2017
Myosin light chain phosphorylation enhances contraction of heart muscle via structural changes in both thick and thin filaments Kampourakis, Thomas; Sun, Yin-Biao; Irving, Malcolm Proceedings of the National Academy of Sciences, Vol. 113, Issue 21 https://doi.org/10.1073/pnas.1602776113	journal	May 2016
Conformational states of smooth muscle myosin. Effects of light chain phosphorylation and ionic strength. Trybus, K. M.; Lowey, S. Journal of Biological Chemistry, Vol. 259, Issue 13 https://doi.org/10.1016/S0021-9258(17)39767-3	journal	July 1984
Hypertrophic and Dilated Cardiomyopathy: Four Decades of Basic Research on Muscle Lead to Potential Therapeutic Approaches to These Devastating Genetic Diseases Spudich, James A. Biophysical Journal, Vol. 106, Issue 6 https://doi.org/10.1016/j.bpj.2014.02.011	journal	March 2014
Biophysical properties of human β-cardiac myosin with converter mutations that cause hypertrophic cardiomyopathy Kawana, Masataka; Sarkar, Saswata S.; Sutton, Shirley Science Advances, Vol. 3, Issue 2 https://doi.org/10.1126/sciadv.1601959	journal	February 2017
Hypertrophic cardiomyopathy and the myosin mesa: viewing an old disease in a new light Trivedi, Darshan V.; Adhikari, Arjun S.; Sarkar, Saswata S. Biophysical Reviews, Vol. 10, Issue 1 https://doi.org/10.1007/s12551-017-0274-6	journal	July 2017
The M·ADP·Pi State Is Required for Helical Order in the Thick Filaments of Skeletal Muscle Xu, S.; Gu, J.; Rhodes, T. Biophysical Journal, Vol. 77, Issue 5 https://doi.org/10.1016/S0006-3495(99)77101-8	journal	November 1999
Regulation of Contraction by the Thick Filaments in Skeletal Muscle Irving, Malcolm Biophysical Journal, Vol. 113, Issue 12 https://doi.org/10.1016/j.bpj.2017.09.037	journal	December 2017
Three-dimensional image reconstruction of dephosphorylated smooth muscle heavy meromyosin reveals asymmetry in the interaction between myosin heads and placement of subfragment 2 Wendt, T.; Taylor, D.; Trybus, K. M. Proceedings of the National Academy of Sciences, Vol. 98, Issue 8 https://doi.org/10.1073/pnas.071051098	journal	April 2001
Myosin ATP turnover rate is a mechanism involved in thermogenesis in resting skeletal muscle fibers Stewart, M. A.; Franks-Skiba, K.; Chen, S. Proceedings of the National Academy of Sciences, Vol. 107, Issue 1 https://doi.org/10.1073/pnas.0909468107	journal	December 2009
The myosin mesa and the basis of hypercontractility caused by hypertrophic cardiomyopathy mutations Nag, Suman; Trivedi, Darshan V.; Sarkar, Saswata S. Nature Structural & Molecular Biology, Vol. 24, Issue 6 https://doi.org/10.1038/nsmb.3408	journal	May 2017
Motion of myosin head domains during activation and force development in skeletal muscle Reconditi, M.; Brunello, E.; Linari, M. Proceedings of the National Academy of Sciences, Vol. 108, Issue 17 https://doi.org/10.1073/pnas.1018330108	journal	April 2011
Effects of hypertrophic and dilated cardiomyopathy mutations on power output by human β-cardiac myosin Spudich, James A.; Aksel, Tural; Bartholomew, Sadie R. The Journal of Experimental Biology, Vol. 219, Issue 2 https://doi.org/10.1242/jeb.125930	journal	January 2016
Myosin filament activation in the heart is tuned to the mechanical task Reconditi, Massimo; Caremani, Marco; Pinzauti, Francesca Proceedings of the National Academy of Sciences, Vol. 114, Issue 12 https://doi.org/10.1073/pnas.1619484114	journal	March 2017
Single-Molecule Mechanics of R403Q Cardiac Myosin Isolated From the Mouse Model of Familial Hypertrophic Cardiomyopathy Tyska, M. J.; Hayes, E.; Giewat, M. Circulation Research, Vol. 86, Issue 7 https://doi.org/10.1161/01.RES.86.7.737	journal	April 2000
Effect of substrate on freeze-dried and shadowed protein structures Trinick, J.; Elliott, A. Journal of Microscopy, Vol. 126, Issue 2 https://doi.org/10.1111/j.1365-2818.1982.tb00364.x	journal	May 1982
A New State of Cardiac Myosin with Very Slow ATP Turnover: A Potential Cardioprotective Mechanism in the Heart Hooijman, Pleuni; Stewart, Melanie A.; Cooke, Roger Biophysical Journal, Vol. 100, Issue 8 https://doi.org/10.1016/j.bpj.2011.02.061	journal	April 2011
The Genetic Basis for Cardiomyopathy Seidman, J. G.; Seidman, Christine Cell, Vol. 104, Issue 4 https://doi.org/10.1016/S0092-8674(01)00242-2	journal	February 2001
Conserved Intramolecular Interactions Maintain Myosin Interacting-Heads Motifs Explaining Tarantula Muscle Super-Relaxed State Structural Basis Alamo, Lorenzo; Qi, Dan; Wriggers, Willy Journal of Molecular Biology, Vol. 428, Issue 6 https://doi.org/10.1016/j.jmb.2016.01.027	journal	March 2016
Mechanisms of the Frank-Starling Law of the Heart: The Beat Goes On John Solaro, R. Biophysical Journal, Vol. 93, Issue 12 https://doi.org/10.1529/biophysj.107.117200	journal	December 2007
Cardiac muscle function across the natural history of a genetic minipig model of hypertrophic cardiomyopathy Henze, Marcus; Anderson, Robert; Wong, Fiona Journal of Molecular and Cellular Cardiology, Vol. 112 https://doi.org/10.1016/j.yjmcc.2017.07.106	journal	November 2017

Similar Records

Myosin dilated cardiomyopathy mutation S532P disrupts actomyosin interactions, leading to altered muscle kinetics, reduced locomotion, and cardiac dilation in Drosophila

Journal Article · Thu Aug 19 00:00:00 EDT 2021 · Molecular Biology of the Cell · OSTI ID:1464331

Trujillo, Adriana S.; Hsu, Karen H.; Puthawala, Joy; +6 more

Molecular basis of force-pCa relation in MYL2 cardiomyopathy mice: Role of the super-relaxed state of myosin

Journal Article · Thu Feb 17 00:00:00 EST 2022 · Proceedings of the National Academy of Sciences of the United States of America · OSTI ID:1464331

Yuan, Chen-Ching; Kazmierczak, Katarzyna; Liang, Jingsheng; +3 more

Homologous mutations in human β, embryonic, and perinatal muscle myosins have divergent effects on molecular power generation

Journal Article · Tue Feb 20 00:00:00 EST 2024 · Proceedings of the National Academy of Sciences of the United States of America · OSTI ID:1464331

Liu, Chao; Karabina, Anastasia; Meller, Artur; +6 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
myosin
super relaxed state
interacting heads motif
cardiac inhibitor
mavacamten

Title: Deciphering the super relaxed state of human β-cardiac myosin and the mode of action of mavacamten from myosin molecules to muscle fibers

Citation Formats

References (64)

Similar Records

Related Subjects