Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization
Ahmet Yildiz,1Joseph N. Forkey,3Sean A. McKinney,1,2Taekjip Ha,1,2Yale E. Goldman,3Paul R. Selvin1,2*
Myosin V is a dimeric molecular motor that moves processivelyon actin, with the center of mass moving 37 nanometers for eachadenosine triphosphate hydrolyzed. We have labeled myosin Vwith a single fluorophore at different positions in the light-chaindomain and measured the step size with a standard deviationof <1.5 nanometers, with 0.5-second temporal resolution,and observation times of minutes. The step size alternates between37 + 2x nm and 37 2x, where x is the distance alongthe direction of motion between the dye and the midpoint betweenthe two heads. These results strongly support a hand-over-handmodel of motility, not an inchworm model.
1 Center for Biophysics and Computational Biology, University of Illinois, Urbana-Champaign, IL 61801, USA. 2 Physics Department, University of Illinois, Urbana-Champaign, IL 61801, USA. 3 University of Pennsylvania, Pennsylvania Muscle Institute, Philadelphia, PA 19104, USA.
* To whom correspondence should be addressed. E-mail: selvin{at}uiuc.edu
Conservation of the regulated structure of folded myosin 2 in species separated by at least 600 million years of independent evolution.
H. S. Jung, S. A. Burgess, N. Billington, M. Colegrave, H. Patel, J. M. Chalovich, P. D. Chantler, and P. J. Knight (2008)
PNAS
105, 6022-6026
|Abstract »|Full Text »|PDF »
The mitotic kinesin CENP-E is a processive transport motor.
H. Yardimci, M. van Duffelen, Y. Mao, S. S. Rosenfeld, and P. R. Selvin (2008)
PNAS
105, 6016-6021
|Abstract »|Full Text »|PDF »
Human Myosin Vc Is a Low Duty Ratio, Nonprocessive Molecular Motor.
Y. Takagi, Y. Yang, I. Fujiwara, D. Jacobs, R. E. Cheney, J. R. Sellers, and M. Kovacs (2008)
J. Biol. Chem.
283, 8527-8537
|Abstract »|Full Text »|PDF »
Agnostic Particle Tracking for Three-Dimensional Motion of Cellular Granules and Membrane-Tethered Bead Dynamics.
K. V. Desai, T. G. Bishop, L. Vicci, E. T. O'Brien Sr., R. M. Taylor 2nd, and R. Superfine (2008)
Biophys. J.
94, 2374-2384
|Abstract »|Full Text »|PDF »
Nanometer-scale mapping and single-molecule detection with color-coded nanoparticle probes.
A. Agrawal, R. Deo, G. D. Wang, M. D. Wang, and S. Nie (2008)
PNAS
105, 3298-3303
|Abstract »|Full Text »|PDF »
Fast Fluorescence Laser Tracking Microrheometry, I: Instrument Development.
M. Jonas, H. Huang, R. D. Kamm, and P. T. C. So (2008)
Biophys. J.
94, 1459-1469
|Abstract »|Full Text »|PDF »
Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy.
Single-molecule tracking of mRNA exiting from RNA polymerase II.
J. Andrecka, R. Lewis, F. Bruckner, E. Lehmann, P. Cramer, and J. Michaelis (2008)
PNAS
105, 135-140
|Abstract »|Full Text »|PDF »
Visualizing the splicing of single pre-mRNA molecules in whole cell extract.
D. J. Crawford, A. A. Hoskins, L. J. Friedman, J. Gelles, and M. J. Moore (2008)
RNA
14, 170-179
|Abstract »|Full Text »|PDF »
Skeletal muscle resists stretch by rapid binding of the second motor domain of myosin to actin.
E. Brunello, M. Reconditi, R. Elangovan, M. Linari, Y.-B. Sun, T. Narayanan, P. Panine, G. Piazzesi, M. Irving, and V. Lombardi (2007)
PNAS
104, 20114-20119
|Abstract »|Full Text »|PDF »
The motor activity of myosin-X promotes actin fiber convergence at the cell periphery to initiate filopodia formation.
Fluorescence Imaging with One-Nanometer Accuracy (FIONA).
P. R. Selvin, T. Lougheed, M. Tonks Hoffman, H. Park, H. Balci, B. H. Blehm, and E. Toprak (2007)
CSH Protocols
2007, pdb.top27
|Abstract »|Full Text »
Myosin V stepping mechanism.
G. Cappello, P. Pierobon, C. Symonds, L. Busoni, J. Christof, M. Gebhardt, M. Rief, and J. Prost (2007)
PNAS
104, 15328-15333
|Abstract »|Full Text »|PDF »
Myo4p is a monomeric myosin with motility uniquely adapted to transport mRNA.
B. D. Dunn, T. Sakamoto, M.-S. S. Hong, J. R. Sellers, and P. A. Takizawa (2007)
J. Cell Biol.
178, 1193-1206
|Abstract »|Full Text »|PDF »
Multicolor Super-Resolution Imaging with Photo-Switchable Fluorescent Probes.
M. Bates, B. Huang, G. T. Dempsey, and X. Zhuang (2007)
Science
317, 1749-1753
|Abstract »|Full Text »|PDF »
Engineering the Processive Run Length of Myosin V.
A. R. Hodges, E. B. Krementsova, and K. M. Trybus (2007)
J. Biol. Chem.
282, 27192-27197
|Abstract »|Full Text »|PDF »
From the Cover: Real-time observations of single bacteriophage {lambda} DNA ejections in vitro.
P. Grayson, L. Han, T. Winther, and R. Phillips (2007)
PNAS
104, 14652-14657
|Abstract »|Full Text »|PDF »
Toward Protein Structure In Situ: Comparison of Two Bifunctional Rhodamine Adducts of Troponin C.
O. Julien, Y.-B. Sun, A. C. Knowles, B. D. Brandmeier, R. E. Dale, D. R. Trentham, J. E. T. Corrie, B. D. Sykes, and M. Irving (2007)
Biophys. J.
93, 1008-1020
|Abstract »|Full Text »|PDF »
Single-molecule Chemistry and Biology Special Feature: New directions in single-molecule imaging and analysis.
Tracking melanosomes inside a cell to study molecular motors and their interaction.
C. Kural, A. S. Serpinskaya, Y.-H. Chou, R. D. Goldman, V. I. Gelfand, and P. R. Selvin (2007)
PNAS
104, 5378-5382
|Abstract »|Full Text »|PDF »
From the Cover: Myosin Va maneuvers through actin intersections and diffuses along microtubules.
M. Y. Ali, E. B. Krementsova, G. G. Kennedy, R. Mahaffy, T. D. Pollard, K. M. Trybus, and D. M. Warshaw (2007)
PNAS
104, 4332-4336
|Abstract »|Full Text »|PDF »
Rapid DNA mapping by fluorescent single molecule detection.
M. Xiao, A. Phong, C. Ha, T.-F. Chan, D. Cai, L. Leung, E. Wan, A. L. Kistler, J. L. DeRisi, P. R. Selvin, et al. (2007)
Nucleic Acids Res.
35, e16
|Abstract »|Full Text »|PDF »
In vivo Real-time Tracking of Single Quantum Dots Conjugated with Monoclonal Anti-HER2 Antibody in Tumors of Mice.
H. Tada, H. Higuchi, T. M. Wanatabe, and N. Ohuchi (2007)
Cancer Res.
67, 1138-1144
|Abstract »|Full Text »|PDF »
The unique insert at the end of the myosin VI motor is the sole determinant of directionality.
H. Park, A. Li, L.-Q. Chen, A. Houdusse, P. R. Selvin, and H. L. Sweeney (2007)
PNAS
104, 778-783
|Abstract »|Full Text »|PDF »
Wide-field subdiffraction imaging by accumulated binding of diffusing probes.
Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy.
S. T. Hess, T. P. K. Girirajan, and M. D. Mason (2006)
Biophys. J.
91, 4258-4272
|Abstract »|Full Text »|PDF »
Extracting Dwell Time Sequences from Processive Molecular Motor Data.
L. S. Milescu, A. Yildiz, P. R. Selvin, and F. Sachs (2006)
Biophys. J.
91, 3135-3150
|Abstract »|Full Text »|PDF »
Regulation of Myosin V Processivity by Calcium at the Single Molecule Level.
H. Lu, E. B. Krementsova, and K. M. Trybus (2006)
J. Biol. Chem.
281, 31987-31994
|Abstract »|Full Text »|PDF »
Analysis Method for Measuring Submicroscopic Distances with Blinking Quantum Dots.
B. C. Lagerholm, L. Averett, G. E. Weinreb, K. Jacobson, and N. L. Thompson (2006)
Biophys. J.
91, 3050-3060
|Abstract »|Full Text »|PDF »
Functional Studies of Individual Myosin Molecules.
J. A DANTZIG, T. Y LIU, and Y. E GOLDMAN (2006)
Ann. N.Y. Acad. Sci.
1080, 1-18
|Abstract »|Full Text »|PDF »
A Kinetic Model Describing the Processivity of Myosin-V.
K. I. Skau, R. B. Hoyle, and M. S. Turner (2006)
Biophys. J.
91, 2475-2489
|Abstract »|Full Text »|PDF »
A Molecular Dynamics Study and Free Energy Analysis of Complexes between the Mlc1p Protein and Two IQ Motif Peptides.
A. Ganoth, R. Friedman, E. Nachliel, and M. Gutman (2006)
Biophys. J.
91, 2436-2450
|Abstract »|Full Text »|PDF »
Spatially and Temporally Synchronized Atomic Force and Total Internal Reflection Fluorescence Microscopy for Imaging and Manipulating Cells and Biomolecules.
M. S. Z. Kellermayer, A. Karsai, A. Kengyel, A. Nagy, P. Bianco, T. Huber, A. Kulcsar, C. Niedetzky, R. Proksch, and L. Grama (2006)
Biophys. J.
91, 2665-2677
|Abstract »|Full Text »|PDF »
Autoinhibition regulates the motility of the C. elegans intraflagellar transport motor OSM-3.
M. Imanishi, N. F. Endres, A. Gennerich, and R. D. Vale (2006)
J. Cell Biol.
174, 931-937
|Abstract »|Full Text »|PDF »
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution.
E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess (2006)
Science
313, 1642-1645
|Abstract »|Full Text »|PDF »
Maximum Likelihood Estimation of Molecular Motor Kinetics from Staircase Dwell-Time Sequences.
L. S. Milescu, A. Yildiz, P. R. Selvin, and F. Sachs (2006)
Biophys. J.
91, 1156-1168
|Abstract »|Full Text »|PDF »
The Globular Tail Domain of Myosin Va Functions as an Inhibitor of the Myosin Va Motor.
X.-d. Li, H. S. Jung, K. Mabuchi, R. Craig, and M. Ikebe (2006)
J. Biol. Chem.
281, 21789-21798
|Abstract »|Full Text »|PDF »
Viewing Dynamic Assembly of Molecular Complexes by Multi-Wavelength Single-Molecule Fluorescence.
J. C. M. Gebhardt, A. E.-M. Clemen, J. Jaud, and M. Rief (2006)
PNAS
103, 8680-8685
|Abstract »|Full Text »|PDF »
Cargo-Binding Makes a Wild-Type Single-Headed Myosin-VI Move Processively.
M. Iwaki, H. Tanaka, A. H. Iwane, E. Katayama, M. Ikebe, and T. Yanagida (2006)
Biophys. J.
90, 3643-3652
|Abstract »|Full Text »|PDF »
Motion Matters: Secretory Granule Motion Adjacent to the Plasma Membrane and Exocytosis.
M. W. Allersma, M. A. Bittner, D. Axelrod, and R. W. Holz (2006)
Mol. Biol. Cell
17, 2424-2438
|Abstract »|Full Text »|PDF »
Defocused orientation and position imaging (DOPI) of myosin V.
E. Toprak, J. Enderlein, S. Syed, S. A. McKinney, R. G. Petschek, T. Ha, Y. E. Goldman, and P. R. Selvin (2006)
PNAS
103, 6495-6499
|Abstract »|Full Text »|PDF »
The fluorescent toolbox for assessing protein location and function..
B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien (2006)
Science
312, 217-224
|Abstract »|Full Text »|PDF »
Direct Observation of Abortive Initiation and Promoter Escape within Single Immobilized Transcription Complexes.
E. Margeat, A. N. Kapanidis, P. Tinnefeld, Y. Wang, J. Mukhopadhyay, R. H. Ebright, and S. Weiss (2006)
Biophys. J.
90, 1419-1431
|Abstract »|Full Text »|PDF »
Multiple events on single molecules: Unbiased estimation in single-molecule biophysics.
Nuclear transport of single molecules: dwell times at the nuclear pore complex.
U. Kubitscheck, D. Grunwald, A. Hoekstra, D. Rohleder, T. Kues, J. P. Siebrasse, and R. Peters (2005)
J. Cell Biol.
168, 233-243
|Abstract »|Full Text »|PDF »
Static and Dynamic Errors in Particle Tracking Microrheology.
Intracellular actin-based transport: How far you go depends on how often you switch.
J. Snider, F. Lin, N. Zahedi, V. Rodionov, C. C. Yu, and S. P. Gross (2004)
PNAS
101, 13204-13209
|Abstract »|Full Text »|PDF »
Myosin VI Steps via a Hand-over-Hand Mechanism with Its Lever Arm Undergoing Fluctuations when Attached to Actin.
A. Yildiz, H. Park, D. Safer, Z. Yang, L.-Q. Chen, P. R. Selvin, and H. L. Sweeney (2004)
J. Biol. Chem.
279, 37223-37226
|Abstract »|Full Text »|PDF »
Nanometer Localization of Single Green Fluorescent Proteins: Evidence that Myosin V Walks Hand-Over-Hand via Telemark Configuration.
G. E. Snyder, T. Sakamoto, J. A. Hammer III, J. R. Sellers, and P. R. Selvin (2004)
Biophys. J.
87, 1776-1783
|Abstract »|Full Text »|PDF »
Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave.
A. Sarkar, R. B. Robertson, and J. M. Fernandez (2004)
PNAS
101, 12882-12886
|Abstract »|Full Text »|PDF »
From the Cover: The dynamics of genomic-length DNA molecules in 100-nm channels.
J. O. Tegenfeldt, C. Prinz, H. Cao, S. Chou, W. W. Reisner, R. Riehn, Y. M. Wang, E. C. Cox, J. C. Sturm, P. Silberzan, et al. (2004)
PNAS
101, 10979-10983
|Abstract »|Full Text »|