Myosin cleft closure determines the energetics of the actomyosin interaction

Formation of the strong binding interaction between actin and myosin is essential for force generation in muscle and in cytoskeletal motor systems. To clarify the role of the closure of myosin's actin‐binding cleft in the actomyosin interaction, we performed rapid kinetic, spectroscopic, and calorimetric experiments and atomic‐level energetic calculations on a variety of myosin isoforms for which atomic structures are available. Surprisingly, we found that the endothermic actin‐binding profile of vertebrate skeletal muscle myosin subfragment‐1 is unique among studied myosins. We show that the diverse propensity of myo‐sins for cleft closure determines different energetic profiles as well as structural and kinetic pathways of actin binding. Depending on the type of myosin, strong actin binding may occur via induced‐fit or conformational preselection mechanisms. However, cleft closure does not directly determine the kinetics and affinity of actin binding. We also show that cleft closure is enthal‐pically unfavorable, reflecting the development of an internal strain within myosin in order to adopt precise steric complementarity to the actin filament. We propose that cleft closure leads to an increase in the torsional strain of myosin's central β‐sheet that has been proposed to serve as an allosteric energy‐transducing spring during force generation.—Takacs, B., O'Neall‐Hennessey, E., Hetenyi, C., Kardos, J., Szent‐Gyorgyi, A. G., Kovacs, M. Myosin cleft closure determines the energetics of the actomyosin interaction. FASEBJ. 25, 111–121 (2011). www.fasebj.org

[1]  Adrian O. Olivares,et al.  Myosin isoform determines the conformational dynamics and cooperativity of actin filaments in the strongly bound actomyosin complex. , 2010, Journal of molecular biology.

[2]  Jianpeng Ma,et al.  CHARMM: The biomolecular simulation program , 2009, J. Comput. Chem..

[3]  I. Rayment,et al.  Actin-binding cleft closure in myosin II probed by site-directed spin labeling and pulsed EPR , 2008, Proceedings of the National Academy of Sciences.

[4]  Martin Karplus,et al.  Allosteric Communication in Myosin V: From Small Conformational Changes to Large Directed Movements , 2008, PLoS Comput. Biol..

[5]  Alfonso T. García-Sosa,et al.  Structure-based calculation of drug efficiency indices , 2007, Bioinform..

[6]  C. Camacho,et al.  FastContact: a free energy scoring tool for protein–protein complex structures , 2007, Nucleic Acids Res..

[7]  C. Cohen,et al.  Rigor-like structures from muscle myosins reveal key mechanical elements in the transduction pathways of this allosteric motor. , 2007, Structure.

[8]  Clive R. Bagshaw,et al.  Reversible movement of switch 1 loop of myosin determines actin interaction , 2007, The EMBO journal.

[9]  Carl A. Morris,et al.  The structure of the myosin VI motor reveals the mechanism of directionality reversal , 2005, Nature.

[10]  M. Konrad,et al.  Kinetic and Thermodynamic Properties of the Ternary Complex between F-actin, Myosin Subfragment 1 and Adenosine 5′-[β,γ-imido]triphosphate , 2005 .

[11]  Chao Zhang,et al.  FastContact: rapid estimate of contact and binding free energies , 2005, Bioinform..

[12]  L. Faller,et al.  Ionic-strength dependence of the conformational change in the unphosphorylated sodium pump. , 2005, Biochemistry.

[13]  Kenneth C Holmes,et al.  The molecular mechanism of muscle contraction. , 2005, Advances in protein chemistry.

[14]  K C Holmes,et al.  The structure of the rigor complex and its implications for the power stroke. , 2004, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[15]  H. Sweeney,et al.  The motor mechanism of myosin V: insights for muscle contraction. , 2004, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[16]  Anne Houdusse,et al.  Three myosin V structures delineate essential features of chemo‐mechanical transduction , 2004, The EMBO journal.

[17]  R. Cooke,et al.  The force exerted by a muscle cross-bridge depends directly on the strength of the actomyosin bond. , 2004, Biophysical journal.

[18]  C. Yengo,et al.  Functional role of loop 2 in myosin V. , 2004, Biochemistry.

[19]  M. Geeves,et al.  A novel stopped-flow method for measuring the affinity of actin for myosin head fragments using μg quantities of protein , 1996, Journal of Muscle Research & Cell Motility.

[20]  D. Manstein,et al.  Overexpression of myosin motor domains in Dictyostelium: screening of transformants and purification of the affinity tagged protein , 1995, Journal of Muscle Research and Cell Motility.

[21]  R. Goody,et al.  Kinetics of acto-S1 interaction as a guide to a model for the crossbridge cycle , 1984, Journal of Muscle Research & Cell Motility.

[22]  T. Pollard,et al.  Pyrene actin: documentation of the validity of a sensitive assay for actin polymerization , 1983, Journal of Muscle Research & Cell Motility.

[23]  Haruki Nakamura,et al.  Announcing the worldwide Protein Data Bank , 2003, Nature Structural Biology.

[24]  J. Sellers,et al.  Functional Divergence of Human Cytoplasmic Myosin II , 2003, Journal of Biological Chemistry.

[25]  Clive R. Bagshaw,et al.  Myosin cleft movement and its coupling to actomyosin dissociation , 2003, Nature Structural Biology.

[26]  Rasmus R. Schröder,et al.  Electron cryo-microscopy shows how strong binding of myosin to actin releases nucleotide , 2003, Nature.

[27]  Carl A. Morris,et al.  A structural state of the myosin V motor without bound nucleotide , 2003, Nature.

[28]  A. Becker,et al.  A structural model for actin-induced nucleotide release in myosin , 2003, Nature Structural Biology.

[29]  Justin E. Molloy,et al.  Neck Length and Processivity of Myosin V* , 2003, Journal of Biological Chemistry.

[30]  J. Sellers,et al.  Kinetic Mechanism of Non-muscle Myosin IIB , 2003, Journal of Biological Chemistry.

[31]  S. Rosenfeld,et al.  Myosin IIB Is Unconventionally Conventional* , 2003, Journal of Biological Chemistry.

[32]  C. Yengo,et al.  Actin-induced Closure of the Actin-binding Cleft of Smooth Muscle Myosin* , 2002, The Journal of Biological Chemistry.

[33]  Clive R. Bagshaw,et al.  Resolution of conformational states of Dictyostelium myosin II motor domain using tryptophan (W501) mutants: implications for the open-closed transition identified by crystallography. , 2000, Biochemistry.

[34]  Amber L. Wells,et al.  The kinetic mechanism of myosin V. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[35]  C. Yengo,et al.  Intrinsic tryptophan fluorescence identifies specific conformational changes at the actomyosin interface upon actin binding and ADP release. , 1999, Biochemistry.

[36]  V. Kalabokis,et al.  Cooperativity and regulation of scallop myosin and myosin fragments. , 1997, Biochemistry.

[37]  D. Manstein,et al.  Dictyostelium discoideum myosin II: characterization of functional myosin motor fragments. , 1997, Biochemistry.

[38]  M. Geeves,et al.  A novel stopped-flow method for measuring the affinity of actin for myosin head fragments using microgram quantities of protein. , 1996, Journal of muscle research and cell motility.

[39]  H. Schuster,et al.  Cloning vectors for the production of proteins in Dictyostelium discoideum. , 1995, Gene.

[40]  D. Manstein,et al.  Kinetic characterization of a cytoplasmic myosin motor domain expressed in Dictyostelium discoideum. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Ivan Rayment,et al.  Three-dimensional atomic model of F-actin decorated with Dictyostelium myosin S1 , 1993, Nature.

[42]  D A Winkelmann,et al.  Three-dimensional structure of myosin subfragment-1: a molecular motor. , 1993, Science.

[43]  E. Taylor.  Kinetic studies on the association and dissociation of myosin subfragment 1 and actin. , 1991, The Journal of biological chemistry.

[44]  M. Geeves,et al.  The effect of nucleotide upon a specific isomerization of actomyosin subfragment 1. , 1988, The Biochemical journal.

[45]  M. Geeves,et al.  Pressure-relaxation studies of pyrene-labelled actin and myosin subfragment 1 from rabbit skeletal muscle. Evidence for two states of acto-subfragment 1. , 1985, The Biochemical journal.

[46]  T Sekine,et al.  A streamlined method of subfragment one preparation from myosin. , 1985, Journal of biochemistry.

[47]  R. Woledge,et al.  Microcalorimetric measurement of the enthalpy of binding of rabbit skeletal myosin subfragment 1 and heavy meromyosin to F-actin. , 1984, The Journal of biological chemistry.

[48]  M. Konrad,et al.  Kinetic and thermodynamic properties of the ternary complex between F-actin, myosin subfragment 1 and adenosine 5'-[beta, gamma-imido]triphosphate. , 1982, European Journal of Biochemistry.

[49]  S. Lowey,et al.  Preparation of myosin and its subfragments from rabbit skeletal muscle. , 1982, Methods in enzymology.

[50]  W. Stafford,et al.  Regulatory properties of single-headed fragments of scallop myosin. , 1979, Biochemistry.

[51]  S. Lowey,et al.  Interaction of myosin subfragments with F-actin. , 1978, Biochemistry.

[52]  S. Highsmith.  The effects of temperature and salts on myosin subfragment-1 and F-actin association. , 1977, Archives of biochemistry and biophysics.

[53]  J. Spudich,et al.  The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. , 1971, The Journal of biological chemistry.