Functional Studies of Individual Myosin Molecules
JODY A. DANTZIG
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authorTIM Y. LIU
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authorYALE E. GOLDMAN
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authorJODY A. DANTZIG
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authorTIM Y. LIU
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authorYALE E. GOLDMAN
University of Pennsylvania School of Medicine, Pennsylvania Muscle Institute, Philadelphia, Pennsylvania 19104-6083, USA
Search for more papers by this authore-mail: [email protected]
Abstract
Abstract: The “conventional” isoform of myosin that polymerizes into filaments (myosin II) is the molecular motor powering contraction in all three types of muscle. Considerable attention has been paid to the developmental progression, isoform distribution, and mutations that affect myocardial development, function, and adaptation. Optical trap (laser tweezer) experiments and various types of high-resolution fluorescence microscopy, capable of interrogating individual protein motors, are revealing novel and detailed information about their functionally relevant nanometer motions and pico-Newton forces. Single-molecule laser tweezer studies of cardiac myosin isoforms and their mutants have helped to elucidate the pathogenesis of familial hypertrophic cardiomyopathies. Surprisingly, some disease mutations seem to enhance myosin function. More broadly, the myosin superfamily includes more than 20 nonfilamentous members with myriad cellular functions, including targeted organelle transport, endocytosis, chemotaxis, cytokinesis, modulation of sensory systems, and signal transduction. Widely varying genetic, developmental and functional disorders of the nervous, pigmentation, and immune systems have been described in accordance with these many roles. Compared to the collective nature of myosin II, some myosin family members operate with only a few partners or even alone. Individual myosin V and VI molecules can carry cellular vesicular cargos much farther distances than their own size. Laser tweezer mechanics, single-molecule fluorescence polarization, and imaging with nanometer precision have elucidated the very different mechano-chemical properties of these isoforms. Critical contributions of nonsarcomeric myosins to myocardial development and adaptation are likely to be discovered in future studies, so these techniques and concepts may become important in cardiovascular research.
REFERENCES
- 1 Barbara, P.F. 2005. Single-molecule spectroscopy. Acc. Chem. Res. 38: 503.
- 2 Rosenberg, S.A., M.E. Quinlan, J.N. Forkey & Y.E. Goldman. 2005. Rotational motions of macro-molecules by single-molecule fluorescence microscopy. Acc. Chem. Res. 38: 583–593.
- 3 Bokinsky, G. & X. Zhuang. 2005. Single-molecule RNA folding. Acc. Chem. Res. 38: 566–573.
- 4 Rasnik, I., S.A. McKinney & T. Ha. 2005. Surfaces and orientations: much to FRET about? Acc. Chem. Res. 38: 542–548.
- 5 Yildiz, A. & P.R. Selvin. 2005. Fluorescence imaging with one nanometer accuracy: application to molecular motors. Acc. Chem. Res. 38: 574–582.
- 6 Ashkin, A. & J.M. Dziedzic. 1987. Optical trapping and manipulation of viruses and bacteria. Science 235: 1517–1520.
- 7 Kuo, S.C. 2001. Using optics to measure biological forces and mechanics. Traffic 2: 757–763.
- 8 Knight, A.E., G. Mashanov & J.E. Molloy. 2005. Single molecule measurements and biological motors. Eur. Biophys. J. 35: 89.
- 9 Svoboda, K. & S.M. Block. 1994. Biological applications of optical forces. Annu. Rev. Biophys. Biomol. Struct. 23: 247–285.
- 10 Sheetz, M.P. 1998. Laser Tweezers in Cell Biology, Vol. 55. Academic Press. San Diego , CA .
- 11 Rock, R.S., S.E. Rice, A.L. Wells, et al . 2001. Myosin VI is a processive motor with a large step size. Proc. Natl. Acad. Sci. USA 98: 13655–13659.
- 12 Finer, J.T., R.M. Simmons & J.A. Spudich. 1994. Single myosin molecule mechanics: piconewton forces and nanometre steps. Nature 368: 113–119.
- 13 Tyska, M.J., E. Hayes, M. Giewat, et al . 2000. Single-molecule mechanics of R403Q cardiac myosin isolated from the mouse model of familial hypertrophic cardiomyopathy. Circ. Res. 86: 737–744.
- 14 Takagi, Y., E.E. Homsher, Y.E. Goldman, et al . 2006. Force generation in single conventional actomyosin complexes under high dynamic load. Biophys. J. 90: 1295–1307.
- 15 Vanzi, F., Y. Takagi, H. Shuman, et al . 2005. Mechanical studies of single ribosome/mRNA complexes. Biophys. J. 89: 1909–1919.
- 16 Seidman J.G. & C. Seidman. 2001. The genetic basis for cardiomyopathy: from mutation identification to mechanistic paradigms. Cell 104: 557–567.
- 17 Geisterfer-Lowrance, A.A.T., S. Kass, G. Tanigawa, et al . 1990. A molecular basis for familial hypertrophic cardiomyopathy: a β cardiac myosin heavy chain gene missense mutation. Cell 62: 999–1006.
- 18 Bement, W.M. & M.S. Mooseker. 1995. TEDS rule: a molecular rationale for differential regulation of myosins by phosphorylation of the heavy chain head. Cell Motil. Cytoskel. 31: 87–92.
- 19 Cuda, G., L. Fananapazir, W.S. Zhu, et al . 1993. Skeletal muscle expression and abnormal function of β-myosin in hypertrophic cardiomyopathy. J. Clin. Invest. 91: 2861–2865.
- 20 Cuda, G., L. Fananapazir, N.D. Epstein, et al . 1997. The in vitro motility activity of β-cardiac myosin depends on the nature of the β-myosin heavy chain gene mutation in hypertrophic cardiomyopathy. J. Musc. Res. Cell Motil. 18: 275–283.
- 21 Sweeney, H.L., A.J. Straceski, L.A. Leinwand, et al . 1994. Heterologous expression of a cardiomyopathic myosin that is defective in its actin interaction. J. Biol. Chem. 269: 1603–1605.
- 22 Sata, M. & M. Ikebe. 1996. Functional analysis of the mutations in the human cardiac β-myosin that are responsible for familial hypertrophic cardiomyopathy. Implication for the clinical outcome. J. Clin. Invest. 98: 2866–2873.
- 23 Roopnarine, O. & L.A. Leinwand. 1998. Functional analysis of myosin mutations that cause familial hypertrophic cardiomyopathy. Biophys. J. 75: 3023–3030.
- 24 Thompson, C.H., G.J. Kemp, D.J. Taylor, et al . 1997. Abnormal skeletal muscle bioenergetics in familial hypertrophic cardiomyopathy. Heart 78: 177–181.
- 25 Blanchard, E., C. Seidman, J.G. Seidman, et al . 1999. Altered crossbridge kinetics in the αMHC403/+ mouse model of familial hypertrophic cardiomyopathy. Circ. Res. 84: 475–483.
- 26 Lankford, E.B., N.D. Epstein, L. Fananapazir, et al . 1995. Abnormal contractile properties of muscle fibers expressing β-myosin heavy chain gene mutations in patients with hypertrophic cardiomyopathy. J. Clin. Invest. 95: 1409–1414.
- 27 Palmiter, K.A., M.J. Tyska, J.R. Haeberle, et al . 2000. R403Q and L908V mutant β-cardiac myosin from patients with familial hypertrophic cardiomyopathy exhibit enhanced mechanical performance at the single molecule level. J Musc. Res. Cell Motil. 21: 609–620.
- 28 Keller, D.I., C. Coirault, T. Rau, et al . 2004. Human homozygous R403W mutant cardiac myosin presents disproportionate enhancement of mechanical and enzymatic properties. J. Mol. Cell Cardiol. 36: 355–362.
- 29 Alpert, N.R., S.A. Mohiddin, D. Tripodi, et al . 2005. Molecular and phenotypic effects of heterozygous, homozygous, and compound heterozygote myosin heavy-chain mutations. Am. J. Physiol. Heart Circ. Physiol. 288: H1097–H1102.
- 30 Köhler J., G. Winkler, I. Schulte, et al . 2002. Mutation of the myosin converter domain alters cross-bridge elasticity. Proc. Natl. Acad. Sci. USA 99: 3557–3562.
- 31 Moss, R.L. & J.S.A. Periera. 2000. Enhanced myosin function due to a point mutation causing a familial hypertrophic cardiomyopathy. Circ. Res. 86: 720–722.
- 32 Ingwall, J.S. & R.G. Weiss. 2004. Is the failing heart energy starved? On using chemical energy to support cardiac function. Circ. Res. 95: 135–145.
- 33 Nyitrai M. & M.A. Geeves. 2004. Adenosine diphosphate and strain sensitivity in myosin motors. Phil. Trans. R. Soc. B. 359: 1867–1877.
- 34 Ahmad, F., J.G. Seidman & C.E. Seidman. 2005. The genetic basis for cardiac remodeling. Annu. Rev. Genomics Hum. Genet. 6: 185–216.
- 35 Berg, J.S., B.C. Powell & R.E. Cheney. 2001. A millennial myosin census. Mol. Biol. Cell 12: 780–794.
- 36 Hodge, T. & M.J.T.V. Cope. 2000. A myosin family tree. J. Cell Sci. 113: 3353–3354.
- 37 Sanger, J.M., B. Mittal, J.S. Dome, et al . 1989. Analysis of cell division using fluorescently labeled actin and myosin in living PtK2 cells. Cell Motil. Cytoskel. 14: 201–219.
- 38 Devreotes, P. & C. Janetopoulos. 2003. Eukaryotic chemotaxis: distinctions between directional sensing and polarization. J. Biol. Chem. 278: 20445–20448.
- 39 Krendel, M. & M. Mooseker. 2005. Myosins: tails (and heads) of functional diversity. Physiology (Bethesda) 20: 239–251.
- 40 De La Cruz, E.M. & E.M. Ostap. 2004. Relating biochemistry and function in the myosin superfamily. Curr. Opin. Cell Biol. 16: 61–67.
- 41 Gillespie, P.G. & J.L. Cyr. 2004. Myosin-1c, the hair cell's adaptation motor. Annu. Rev. Physiol. 66: 521–545.
- 42 Lister I., S. Schmitz, M. Walker, et al . 2004. A monomeric myosin VI with a large working stroke. EMBO J. 23: 1729–1738.
- 43 Park, H., B. Ramamurthy, M. Travaglia, et al . 2006. Full-length myosin VI dimerizes and moves processively along actin filaments upon monomer clustering. Mol. Cell. 21: 331–336.
- 44 Wells, A.L., A.W. Lin, L.-Q. Chen, et al . 1999. Myosin VI is an actin-based motor that moves backwards. Nature 401: 505–508.
- 45 Altman, D., H.L. Sweeney & J.A. Spudich. 2004. The mechanism of myosin VI translocation and its load-induced anchoring. Cell 116: 737–749.
- 46 Ménétrey, J., A. Bahloul, A.L. Wells, et al . 2005. The structure of the myosin VI motor reveals the mechanism of directionality reversal. Nature 435: 779–785.
- 47 Sun, Y., H.W. Schroeder, J.F. Beausang, et al . 2005. Single molecule fluorescence polarization of calmodulin in myosin VI. Biophys. J. 90: 431a.
- 48 Schuler, G.D., M.S. Boguski, E.A. Stewart, et al . 1996. A gene map of the human genome. Science 274: 540–546.
- 49 Pontius, J.U., L. Wagner & G.D. Schuler. 2003. UniGene: a unified view of the transcriptome. In The NCBI Handbook. National Center for Biotechnology Information. Bethesda , MD . (http://www.ncbi.nlm.nih.gov/UniGene).
- 50 Bouck, J., W. Yu, R. Gibbs & K. Worley. 1999. Comparison of gene indexing databases. Trends Gen. 15: 159–162.
- 51 Burke, J., H. Wang, W. Hide, et al . 1998. Alternative gene form discovery and candidate gene selection from gene indexing projects. Genome Res. 8: 276–290.
- 52 Pandey, A. & F. Lewitter. 1999. Nucleotide sequence databases: a gold mine for biologists. Trends Biochem. Sci. 24: 276–280.
- 53 Yildiz, A., J.N. Forkey, S.A. McKinney, et al . 2003. Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization. Science 300: 2061–2065.
- 54 Yildiz, A., M. Tomishige, R.D. Vale, et al . 2004. Kinesin walks hand-over-hand. Science 303: 676–678.
- 55 Yildiz, A., H. Park, D. Safer, et al . 2004. Myosin VI steps via a hand-over-hand mechanism with its lever arm undergoing fluctuations when attached to actin. J. Biol. Chem. 279: 37223–37226.
- 56 Heuser, J.E. & M.W. Kirschner. 1980. Filament organization revealed in platinum replicas of freeze-dried cytoskeletons. J. Cell Biol. 86: 212–234.
- 57 Medalia, O., I. Weber, A.S. Frangakis, et al . 2002. Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 298: 1209–1213.
- 58 Langford, G.M. 2002. Myosin-V, a versatile motor for short-range vesicle transport. Traffic 3: 859–865.
- 59 Frank, D.J., T. Noguchi & K.G. Miller. 2004. Myosin VI: a structural role in actin organization important for protein and organelle localization and trafficking. Curr. Opin. Cell Biol. 16: 189–194.
- 60 Forkey, J.N., M.E. Quinlan, M.A. Shaw, et al . 2003. Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization. Nature 422: 399–404.
- 61 Quinlan, M.E., J.N. Forkey & Y.E. Goldman. 2005. Orientation of the myosin light chain region by single molecule total internal reflection fluorescence polarization microscopy. Biophys. J. 89: 1132–1142.
- 62 Huxley, A.F. & R. Niedergerke. 1954. Structural changes in muscle during contraction; interference microscopy of living muscle fibres. Nature 173: 971–973.
- 63 Huxley, H. & J. Hanson. 1954. Changes in the cross-striations of muscle during contraction and stretch and their structural interpretation. Nature 173: 973–976.
- 64 Hanson, J. & H.E. Huxley. 1955. The structural basis of contraction in striated muscle. Symp. Soc. Exp. Biol. 9: 228–264.
- 65 Huxley, A.F. 1957. Muscle structure and theories of contraction. Prog. Biophys. Biophys. Chem. 7: 255–318.
- 66 Huxley, H.E. 1969. The mechanism of muscular contraction. Science 164: 1356–1366.
- 67 Huxley, A.F. & R.M. Simmons. 1971. Proposed mechanism of force generation in striated muscle. Nature 233: 533–538.
- 68 Rayment, I., H.M. Holden, M. Whittaker, et al . 1993. Structure of the actin-myosin complex and its implications for muscle contraction. Science 261: 58–65.
- 69 Warshaw, D.M. 2004. Lever arms and necks: a common mechanistic theme across the myosin superfamily. J. Musc. Res. Cell Motil. 25: 467–474.
- 70 Hopkins, S.C., C. Sabido-David, J.E.T. Corrie, et al . 1998. Fluorescence polarization transients from rhodamine isomers on the myosin regulatory light chain in skeletal muscle fibers. Biophys. J. 74: 3093–3110.
- 71 Pollard, T.D. & W.C. Earnshaw. 2002. Cell Biology. Chaps: 35–42. Saunders. Philadelphia , PA .
- 72 Hirokawa, N. & R. Takemura. 2004. Kinesin superfamily proteins and their various functions and dynamics. Exp. Cell Res. 301: 50–59.
- 73 Holzbaur, E.L.F. 2004. Motor neurons rely on motor proteins. Trends Cell Biol. 14: 233–240.
- 74 Brown, M.E. & P.C. Bridgman. 2003. Myosin function in nervous and sensory systems. J. Neurobiol. 58: 118–130.
- 75 Levin, M. 2004. The embryonic origins of left-right asymmetry. Crit. Rev. Oral. Biol. Med. 15: 197–206.
- 76 Qiu, D., S.M. Cheng, L. Wozniak, et al . 2005. Localization and loss-of-function implicates ciliary proteins in early, cytoplasmic roles in left-right asymmetry. Dev. Dyn. 234: 176–189.
- 77 Abel, E.D., H.C. Kaulbach, R. Tian, et al . 1999. Cardiac hypertrophy with preserved contractile function after selective deletion of GLUT4 from the heart. J. Clin. Invest. 104: 1703–1714.
- 78 Avraham, K.B., T. Hasson, T. Sobe, et al . 1997. Characterization of unconventional MYO6, the human homologue of the gene responsible for deafness in Snell's waltzer mice. Hum. Mol. Genet. 6: 1225–1231.
- 79 Mohiddin, S.A., Z.M. Ahmed, A.J. Griffith, et al . 2004. Novel association of hypertrophic cardiomyopathy, sensorineural deafness, and a mutation in unconventional myosin VI (MYO6). J. Med. Genet. 41: 309–314.
- 80 Melchionda, S., N. Ahituv, L. Bisceglia, et al . 2001. MYO6, the human homologue of the gene responsible for deafness in Snell's Waltzer mice, is mutated in autosomal dominant nonsyndromic hearing loss. Am. J. Hum. Genet. 69: 635–640.
- 81 Veugelers, M., M. Bressan, D.A. McDermott, et al . 2004. Mutation of perinatal myosin heavy chain with a Carney complex variant. N. Engl. J. Med. 351: 460–469.
- 82 Pollard, T.D. 2003. The cytoskeleton, cellular motility and the reductionist agenda. Nature 422: 741–745.
- 83 Schliwa, M. & G. Woehlke. 2003. Molecular motors. Nature 422: 759–765.