Letter pubs.acs.org/NanoLett
Local Heat Activation of Single Myosins Based on Optical Trapping of Gold Nanoparticles Mitsuhiro Iwaki,*,†,‡ Atsuko H. Iwane,†,‡ Keigo Ikezaki,§ and Toshio Yanagida†,‡,∥ †
Quantitative Biology Center, RIKEN, Suita, Osaka 5650874, Japan Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 5650874, Japan § School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, 2778561, Japan ∥ Center for Information and Neural Networks, NICT, Suita, Osaka, 5650874, Japan ‡
S Supporting Information *
ABSTRACT: Myosin is a mechano-enzyme that hydrolyzes ATP in order to move unidirectionally along actin filaments. Here we show by single molecule imaging that myosin V motion can be activated by local heat. We constructed a dark-field microscopy that included optical tweezers to monitor 80 nm gold nanoparticles (GNP) bound to single myosin V molecules with nanometer and submillisecond accuracy. We observed 34 nm processive steps along actin filaments like those seen when using 200 nm polystyrene beads (PB) but dwell times (ATPase activity) that were 4.5 times faster. Further, by using DNA nanotechnology (DNA origami) and myosin V as a nanometric thermometer, the temperature gradient surrounding optically trapped GNP could be estimated with nanometer accuracy. We propose our single molecule measurement system should advance quantitative analysis of the thermal control of biological and artificial systems like nanoscale thermal ratchet motors. KEYWORDS: Motor protein, optical trap, single molecule, gold nanoparticle, local heating
T
hydrolyzes ATP, which allows myosin to move unidirectionally along actin filaments by a hand-overhand mechanism.9 In this Letter, we demonstrate by single molecule analysis the mechanical properties of myosin V when activated by the local heating of optically trapped GNP. Relative to myosin V attached to polystyrene beads (PB), we found the stepping rate of myosin V attached to GNP was accelerated 4.5-fold when the enzymatic domain (motor domain) was ∼35 nm from the gold surface and 1.2-fold when ∼170 nm from the surface. At the same time, heating caused the maximum force of myosin V attached to GNP to be about half that when attached to PB. These results show that optically trapped GNP could thermally activate myosin within a few hundred nanometers. We constructed myosin V dimers tagged with Halo-tag15 at the tail end. The Halo-tag was then linked to a Cy3 streptavidin via a biotinylated ligand, which was linked to an 80 nm GNP via a biotinylated alkanethiol self-assembled monolayer (SAM;
emperature modulates many biological phenomena such as enzymatic reaction rates and the opening/closing of membrane channels.1 Recently, it was found that the temperature in a living cell is heterogeneous2−4 due to the local heat of reactions inside organelles such as the nucleus and mitochondria. It has since been proposed that this heterogeneity regulates the activities of intracellular biomolecules and their corresponding functions.2 Indeed, local heating by artificial means, such as gold nanoparticles (GNP), has proven effective in photothermal cancer therapy,5 the remote control of neurons in Caenorhabditis elegans,6 vesicle transport,4 and the contraction of cardiomyocytes.7 However, details on how local heating affects biological function, especially at the single molecule level, are still lacking. Myosin V is a well-characterized motor protein in terms of its single molecule mechanical properties,8−10 atomic structure,11 enzyme properties,12 and physiological function.13 Its structure is composed of a motor domain (or head domain), rodlike domain, which includes a lever-arm 26 nm long,14 and a tail domain, which is responsible for dimerization. The head © XXXX American Chemical Society
Received: December 21, 2014 Revised: February 23, 2015
A
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
Letter
Nano Letters
Figure 1. Experimental design. Myosin V was attached to GNP (80 nm in diameter) via a biotinylated self-assembled monolayer (SAM) of alkanethiol (∼2 nm), fluorescent dye (Cy3)-labeled streptavidin (∼5 nm), and a biotinylated Halo tag (∼3 nm). In total, the distance between the GNP surface and myosin head was ∼35 nm. GNPs were optically trapped by a 1064 nm-focused laser at 120 mW. A fluorescent actin filament was stiffly adsorbed onto a cover glass by an actin binding protein, α actinin.
Figure 1, see Materials and Methods, Supporting Information) or 200 nm fluorescent PB. These fluorescent particles were optically trapped in solution and moved onto a single actin filament adsorbed to the cover glass. Dark-field images of the optically trapped particles were expanded and projected onto a quadrant photodiode to detect displacement with nanometer and submillisecond resolution (Supporting Information Figure S1). When GNP or PB was mixed with myosin V at a stoichiometry of 1:1 pN),8 we only analyzed dwell times at loads lower than 1 pN. (c) Maximum force. Distributions were fit to Gaussians with peaks at 2.3 ± 0.23 pN for PB and 0.92 ± 0.24 pN for GNP. Maximum force was defined as the force just before detachment.
surface (Figure 5a), whereas dwell times varied with distance (Figure 5b): 38 ms for ∼90 nm and 58 ms for ∼170 nm. This latter value is similar to the 72 ms seen when using 200 nm PB, implying the temperature at 170 nm mimics bulk water temperature. This conclusion is based on an estimate for the heat production of an optically trapped PB, which was 1−8 K/ W,25,26 leading to a temperature increase of only 100 nm in diameter) at higher laser power. Maximum force was less when measured at ∼35 nm separation from the optically trapped GNP surface (Figure 3c). The reason can be explained by the force generation mechanism of myosin V. It has been clarified that during force generation, the rear myosin head in the dimer detaches from actin and the front head strongly binds, possibly by taking the ADP bound state (ref 10 and Supporting Information Figure S6). We showed local heating accelerated the ADP release from 72 ms in the PB assay to 15.8 ms in the GNP assay, meaning detachment of the front head from actin is promoted by attaching ATP to the head. This outcome would reduce the possibility of completing force generation and explain the decreased maximum force. Actually, maximum force when measured at ∼90 and 170 nm separation from the optically trapped GNP surface was similar to the PB assay (Supporting Information Figure S4). Rapid ADP release is typical in muscle myosin (myosin II). In muscle, several hundreds of myosin II self-assemble to form myosin filaments and generate force cooperatively.30 Rapid detachment of myosin heads from actin by fast ADP release reduces molecular friction to generate efficient and rapid muscle contraction. Myosin V on the other hand functions as an organelle transporter in cells. Such organelles (mitochondria, and so forth) can have relatively higher temperatures than the surrounding cytosol,2,3 much like our GNP. This local heat could then also be advantageous for rapid and efficient transport by multiple myosin V molecules. To conclude, we constructed an observation system to monitor single myosin dynamics under local heating with high resolution. The mechanical behavior reported the temperature around optically trapped GNPs and its effect on myosin V. Further, by using DNA origami rod and myosin V as a nanometric thermometer, the temperature gradient surrounding optically trapped GNP could be estimated. This method is applicable to other enzyme-based systems like gene expression by RNA polymerase to activate the production of mRNA and to artificial nanoscale systems like thermal ratchet type transport systems, where locally heated Brownian nanoparticles are transported unidirectionally in an asymmetric and periodic potential field.31
■
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS We thank colleagues of the Riken Quantitative Biology Center for discussions and Peter Karagianins for reading the manuscript. This work was supported by Grant-in-Aid for Scientific Research on Innovative Areas (MEXT), Grant-inAids for Young Scientists (B) (JSPS).
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ABBREVIATIONS ATP, Adenosine Triphosphate; ADP, Adenosine Diphosphate; Pi, inorganic Phosphate; GNP, gold nanoparticle; PB, polystyrene bead; SAM, self-assembled monolayer
■
ASSOCIATED CONTENT
S Supporting Information *
Detailed materials and methods and supplementary figures. This material is available free of charge via the Internet at http://pubs.acs.org.
■
REFERENCES
(1) Clapham, D. E. Nature 2003, 426, 517−24. (2) Okabe, K.; Inada, N.; Gota, C.; Harada, Y.; Funatsu, T.; Uchiyama, S. Nat. Commun. 2012, 3, 705. (3) Kiyonaka, S.; Kajimoto, T.; Sakaguchi, R.; Shinmi, D.; OmatsuKanbe, M.; Matsuura, H.; Imamura, H.; Yoshizaki, T.; Hamachi, I.; Morii, T.; Mori, Y. Nat. Methods 2013, 10, 1232−8. (4) Oyama, K.; Takabayashi, M.; Takei, Y.; Arai, S.; Takeoka, S.; Ishiwata, S.; Suzuki, M. Lab Chip 2012, 12, 1591−3. (5) Gobin, A. M.; Lee, M. H.; Halas, N. J.; James, W. D.; Drezek, R. A.; West, J. L. Nano Lett. 2007, 7, 1929−34. (6) Huang, H.; Delikanli, S.; Zeng, H.; Ferkey, D. M.; Pralle, A. Nat. Nanotechnol 2010, 5, 602−6. (7) Oyama, K.; Mizuno, A.; Shintani, S. A.; Itoh, H.; Serizawa, T.; Fukuda, N.; Suzuki, M.; Ishiwata, S. Biochem. Biophys. Res. Commun. 2012, 417, 607−12. (8) Mehta, A. D.; Rock, R. S.; Rief, M.; Spudich, J. A.; Mooseker, M. S.; Cheney, R. E. Nature 1999, 400, 590−3. (9) Yildiz, A.; Forkey, J. N.; McKinney, S. A.; Ha, T.; Goldman, Y. E.; Selvin, P. R. Science 2003, 300, 2061−5. (10) Fujita, K.; Iwaki, M.; Iwane, A. H.; Marcucci, L.; Yanagida, T. Nat. Commun. 2012, 3, 956. (11) Coureux, P. D.; Wells, A. L.; Menetrey, J.; Yengo, C. M.; Morris, C. A.; Sweeney, H. L.; Houdusse, A. Nature 2003, 425, 419−23. (12) De La Cruz, E. M.; Wells, A. L.; Rosenfeld, S. S.; Ostap, E. M.; Sweeney, H. L. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 13726−31. (13) Sellers, J. R.; Weisman, L. S. Myosins A Superfamily of Molecular Motors; Springer: Netherlands, 2008. (14) Vilfan, A. Biophys. J. 2005, 88, 3792−805. (15) Los, G. V.; Encell, L. P.; McDougall, M. G.; Hartzell, D. D.; Karassina, N.; Zimprich, C.; Wood, M. G.; Learish, R.; Ohana, R. F.; Urh, M.; Simpson, D.; Mendez, J.; Zimmerman, K.; Otto, P.; Vidugiris, G.; Zhu, J.; Darzins, A.; Klaubert, D. H.; Bulleit, R. F.; Wood, K. V. ACS Chem. Biol. 2008, 3, 373−82. (16) Rief, M.; Rock, R. S.; Mehta, A. D.; Mooseker, M. S.; Cheney, R. E.; Spudich, J. A. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 9482−6. (17) Kyrsting, A.; Bendix, P. M.; Stamou, D. G.; Oddershede, L. B. Nano Lett. 2011, 11, 888−92. (18) Bendix, P. M.; Reihani, S. N.; Oddershede, L. B. ACS Nano 2010, 4, 2256−62. (19) Seol, Y.; Carpenter, A. E.; Perkins, T. T. Opt. Lett. 2006, 31, 2429−31. (20) Urban, A. S.; Fedoruk, M.; Horton, M. R.; Radler, J. O.; Stefani, F. D.; Feldmann, J. Nano Lett. 2009, 9, 2903−8. (21) Rothemund, P. W. Nature 2006, 440, 297−302. (22) Douglas, S. M.; Dietz, H.; Liedl, T.; Hogberg, B.; Graf, F.; Shih, W. M. Nature 2009, 459, 414−8. (23) Derr, N. D.; Goodman, B. S.; Jungmann, R.; Leschziner, A. E.; Shih, W. M.; Reck-Peterson, S. L. Science 2012, 338, 662−5.
AUTHOR INFORMATION
Corresponding Author
*E-mail: [email protected]. Fax: +81661550112. Author Contributions
M.I. conceived the experiment, constructed the microscope, DNA rod, and streptavidin-coated GNP, carried out the experiments, analyzed the data, and wrote the manuscript. A.H.I. and K.I. constructed and purified the myosin V E
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
Letter
Nano Letters (24) Crivat, G.; Taraska, J. W. Trends Biotechnol. 2012, 30, 8−16. (25) Peterman, E. J.; Gittes, F.; Schmidt, C. F. Biophys. J. 2003, 84, 1308−16. (26) Block, S. M. Optical Tweezers: A New Tool for Biophysics. In Noninvasive Techniques in Cell Biology; Wiley: New York, 1990. (27) Robblee, J. P.; Cao, W.; Henn, A.; Hannemann, D. E.; De La Cruz, E. M. Biochemistry 2005, 44, 10238−49. (28) Savin, A. V.; Mazo, M. A.; Kikot, I. P.; Manevitch, L. I.; Onufriev, A. V. Phys. Rev. B: Condens. Matter 2010, 83, 245406. (29) Takakura, Y.; Tsunashima, M.; Suzuki, J.; Usami, S.; Kakuta, Y.; Okino, N.; Ito, M.; Yamamoto, T. FEBS J. 2009, 276, 1383−97. (30) Phillips, R.; Kondev, J.; Theriot, J. Physical Biology of the Cell; Garland Science: New York, 2009. (31) Feynman, R. P. The Feynman Lectures on Physics; AddisonWesley: Reading, MA, 1963; Vol. 1. (32) Kerssemakers, J. W.; Munteanu, E. L.; Laan, L.; Noetzel, T. L.; Janson, M. E.; Dogterom, M. Nature 2006, 442, 709−12.
F
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
Local Heat Activation of Single Myosins Based on Optical Trapping of Gold Nanoparticles Mitsuhiro Iwaki,*,†,‡ Atsuko H. Iwane,†,‡ Keigo Ikezaki,§ and Toshio Yanagida†,‡,∥ †
Quantitative Biology Center, RIKEN, Suita, Osaka 5650874, Japan Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 5650874, Japan § School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, 2778561, Japan ∥ Center for Information and Neural Networks, NICT, Suita, Osaka, 5650874, Japan ‡
S Supporting Information *
ABSTRACT: Myosin is a mechano-enzyme that hydrolyzes ATP in order to move unidirectionally along actin filaments. Here we show by single molecule imaging that myosin V motion can be activated by local heat. We constructed a dark-field microscopy that included optical tweezers to monitor 80 nm gold nanoparticles (GNP) bound to single myosin V molecules with nanometer and submillisecond accuracy. We observed 34 nm processive steps along actin filaments like those seen when using 200 nm polystyrene beads (PB) but dwell times (ATPase activity) that were 4.5 times faster. Further, by using DNA nanotechnology (DNA origami) and myosin V as a nanometric thermometer, the temperature gradient surrounding optically trapped GNP could be estimated with nanometer accuracy. We propose our single molecule measurement system should advance quantitative analysis of the thermal control of biological and artificial systems like nanoscale thermal ratchet motors. KEYWORDS: Motor protein, optical trap, single molecule, gold nanoparticle, local heating
T
hydrolyzes ATP, which allows myosin to move unidirectionally along actin filaments by a hand-overhand mechanism.9 In this Letter, we demonstrate by single molecule analysis the mechanical properties of myosin V when activated by the local heating of optically trapped GNP. Relative to myosin V attached to polystyrene beads (PB), we found the stepping rate of myosin V attached to GNP was accelerated 4.5-fold when the enzymatic domain (motor domain) was ∼35 nm from the gold surface and 1.2-fold when ∼170 nm from the surface. At the same time, heating caused the maximum force of myosin V attached to GNP to be about half that when attached to PB. These results show that optically trapped GNP could thermally activate myosin within a few hundred nanometers. We constructed myosin V dimers tagged with Halo-tag15 at the tail end. The Halo-tag was then linked to a Cy3 streptavidin via a biotinylated ligand, which was linked to an 80 nm GNP via a biotinylated alkanethiol self-assembled monolayer (SAM;
emperature modulates many biological phenomena such as enzymatic reaction rates and the opening/closing of membrane channels.1 Recently, it was found that the temperature in a living cell is heterogeneous2−4 due to the local heat of reactions inside organelles such as the nucleus and mitochondria. It has since been proposed that this heterogeneity regulates the activities of intracellular biomolecules and their corresponding functions.2 Indeed, local heating by artificial means, such as gold nanoparticles (GNP), has proven effective in photothermal cancer therapy,5 the remote control of neurons in Caenorhabditis elegans,6 vesicle transport,4 and the contraction of cardiomyocytes.7 However, details on how local heating affects biological function, especially at the single molecule level, are still lacking. Myosin V is a well-characterized motor protein in terms of its single molecule mechanical properties,8−10 atomic structure,11 enzyme properties,12 and physiological function.13 Its structure is composed of a motor domain (or head domain), rodlike domain, which includes a lever-arm 26 nm long,14 and a tail domain, which is responsible for dimerization. The head © XXXX American Chemical Society
Received: December 21, 2014 Revised: February 23, 2015
A
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
Letter
Nano Letters
Figure 1. Experimental design. Myosin V was attached to GNP (80 nm in diameter) via a biotinylated self-assembled monolayer (SAM) of alkanethiol (∼2 nm), fluorescent dye (Cy3)-labeled streptavidin (∼5 nm), and a biotinylated Halo tag (∼3 nm). In total, the distance between the GNP surface and myosin head was ∼35 nm. GNPs were optically trapped by a 1064 nm-focused laser at 120 mW. A fluorescent actin filament was stiffly adsorbed onto a cover glass by an actin binding protein, α actinin.
Figure 1, see Materials and Methods, Supporting Information) or 200 nm fluorescent PB. These fluorescent particles were optically trapped in solution and moved onto a single actin filament adsorbed to the cover glass. Dark-field images of the optically trapped particles were expanded and projected onto a quadrant photodiode to detect displacement with nanometer and submillisecond resolution (Supporting Information Figure S1). When GNP or PB was mixed with myosin V at a stoichiometry of 1:1 pN),8 we only analyzed dwell times at loads lower than 1 pN. (c) Maximum force. Distributions were fit to Gaussians with peaks at 2.3 ± 0.23 pN for PB and 0.92 ± 0.24 pN for GNP. Maximum force was defined as the force just before detachment.
surface (Figure 5a), whereas dwell times varied with distance (Figure 5b): 38 ms for ∼90 nm and 58 ms for ∼170 nm. This latter value is similar to the 72 ms seen when using 200 nm PB, implying the temperature at 170 nm mimics bulk water temperature. This conclusion is based on an estimate for the heat production of an optically trapped PB, which was 1−8 K/ W,25,26 leading to a temperature increase of only 100 nm in diameter) at higher laser power. Maximum force was less when measured at ∼35 nm separation from the optically trapped GNP surface (Figure 3c). The reason can be explained by the force generation mechanism of myosin V. It has been clarified that during force generation, the rear myosin head in the dimer detaches from actin and the front head strongly binds, possibly by taking the ADP bound state (ref 10 and Supporting Information Figure S6). We showed local heating accelerated the ADP release from 72 ms in the PB assay to 15.8 ms in the GNP assay, meaning detachment of the front head from actin is promoted by attaching ATP to the head. This outcome would reduce the possibility of completing force generation and explain the decreased maximum force. Actually, maximum force when measured at ∼90 and 170 nm separation from the optically trapped GNP surface was similar to the PB assay (Supporting Information Figure S4). Rapid ADP release is typical in muscle myosin (myosin II). In muscle, several hundreds of myosin II self-assemble to form myosin filaments and generate force cooperatively.30 Rapid detachment of myosin heads from actin by fast ADP release reduces molecular friction to generate efficient and rapid muscle contraction. Myosin V on the other hand functions as an organelle transporter in cells. Such organelles (mitochondria, and so forth) can have relatively higher temperatures than the surrounding cytosol,2,3 much like our GNP. This local heat could then also be advantageous for rapid and efficient transport by multiple myosin V molecules. To conclude, we constructed an observation system to monitor single myosin dynamics under local heating with high resolution. The mechanical behavior reported the temperature around optically trapped GNPs and its effect on myosin V. Further, by using DNA origami rod and myosin V as a nanometric thermometer, the temperature gradient surrounding optically trapped GNP could be estimated. This method is applicable to other enzyme-based systems like gene expression by RNA polymerase to activate the production of mRNA and to artificial nanoscale systems like thermal ratchet type transport systems, where locally heated Brownian nanoparticles are transported unidirectionally in an asymmetric and periodic potential field.31
■
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS We thank colleagues of the Riken Quantitative Biology Center for discussions and Peter Karagianins for reading the manuscript. This work was supported by Grant-in-Aid for Scientific Research on Innovative Areas (MEXT), Grant-inAids for Young Scientists (B) (JSPS).
■
ABBREVIATIONS ATP, Adenosine Triphosphate; ADP, Adenosine Diphosphate; Pi, inorganic Phosphate; GNP, gold nanoparticle; PB, polystyrene bead; SAM, self-assembled monolayer
■
ASSOCIATED CONTENT
S Supporting Information *
Detailed materials and methods and supplementary figures. This material is available free of charge via the Internet at http://pubs.acs.org.
■
REFERENCES
(1) Clapham, D. E. Nature 2003, 426, 517−24. (2) Okabe, K.; Inada, N.; Gota, C.; Harada, Y.; Funatsu, T.; Uchiyama, S. Nat. Commun. 2012, 3, 705. (3) Kiyonaka, S.; Kajimoto, T.; Sakaguchi, R.; Shinmi, D.; OmatsuKanbe, M.; Matsuura, H.; Imamura, H.; Yoshizaki, T.; Hamachi, I.; Morii, T.; Mori, Y. Nat. Methods 2013, 10, 1232−8. (4) Oyama, K.; Takabayashi, M.; Takei, Y.; Arai, S.; Takeoka, S.; Ishiwata, S.; Suzuki, M. Lab Chip 2012, 12, 1591−3. (5) Gobin, A. M.; Lee, M. H.; Halas, N. J.; James, W. D.; Drezek, R. A.; West, J. L. Nano Lett. 2007, 7, 1929−34. (6) Huang, H.; Delikanli, S.; Zeng, H.; Ferkey, D. M.; Pralle, A. Nat. Nanotechnol 2010, 5, 602−6. (7) Oyama, K.; Mizuno, A.; Shintani, S. A.; Itoh, H.; Serizawa, T.; Fukuda, N.; Suzuki, M.; Ishiwata, S. Biochem. Biophys. Res. Commun. 2012, 417, 607−12. (8) Mehta, A. D.; Rock, R. S.; Rief, M.; Spudich, J. A.; Mooseker, M. S.; Cheney, R. E. Nature 1999, 400, 590−3. (9) Yildiz, A.; Forkey, J. N.; McKinney, S. A.; Ha, T.; Goldman, Y. E.; Selvin, P. R. Science 2003, 300, 2061−5. (10) Fujita, K.; Iwaki, M.; Iwane, A. H.; Marcucci, L.; Yanagida, T. Nat. Commun. 2012, 3, 956. (11) Coureux, P. D.; Wells, A. L.; Menetrey, J.; Yengo, C. M.; Morris, C. A.; Sweeney, H. L.; Houdusse, A. Nature 2003, 425, 419−23. (12) De La Cruz, E. M.; Wells, A. L.; Rosenfeld, S. S.; Ostap, E. M.; Sweeney, H. L. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 13726−31. (13) Sellers, J. R.; Weisman, L. S. Myosins A Superfamily of Molecular Motors; Springer: Netherlands, 2008. (14) Vilfan, A. Biophys. J. 2005, 88, 3792−805. (15) Los, G. V.; Encell, L. P.; McDougall, M. G.; Hartzell, D. D.; Karassina, N.; Zimprich, C.; Wood, M. G.; Learish, R.; Ohana, R. F.; Urh, M.; Simpson, D.; Mendez, J.; Zimmerman, K.; Otto, P.; Vidugiris, G.; Zhu, J.; Darzins, A.; Klaubert, D. H.; Bulleit, R. F.; Wood, K. V. ACS Chem. Biol. 2008, 3, 373−82. (16) Rief, M.; Rock, R. S.; Mehta, A. D.; Mooseker, M. S.; Cheney, R. E.; Spudich, J. A. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 9482−6. (17) Kyrsting, A.; Bendix, P. M.; Stamou, D. G.; Oddershede, L. B. Nano Lett. 2011, 11, 888−92. (18) Bendix, P. M.; Reihani, S. N.; Oddershede, L. B. ACS Nano 2010, 4, 2256−62. (19) Seol, Y.; Carpenter, A. E.; Perkins, T. T. Opt. Lett. 2006, 31, 2429−31. (20) Urban, A. S.; Fedoruk, M.; Horton, M. R.; Radler, J. O.; Stefani, F. D.; Feldmann, J. Nano Lett. 2009, 9, 2903−8. (21) Rothemund, P. W. Nature 2006, 440, 297−302. (22) Douglas, S. M.; Dietz, H.; Liedl, T.; Hogberg, B.; Graf, F.; Shih, W. M. Nature 2009, 459, 414−8. (23) Derr, N. D.; Goodman, B. S.; Jungmann, R.; Leschziner, A. E.; Shih, W. M.; Reck-Peterson, S. L. Science 2012, 338, 662−5.
AUTHOR INFORMATION
Corresponding Author
*E-mail: [email protected]. Fax: +81661550112. Author Contributions
M.I. conceived the experiment, constructed the microscope, DNA rod, and streptavidin-coated GNP, carried out the experiments, analyzed the data, and wrote the manuscript. A.H.I. and K.I. constructed and purified the myosin V E
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
Letter
Nano Letters (24) Crivat, G.; Taraska, J. W. Trends Biotechnol. 2012, 30, 8−16. (25) Peterman, E. J.; Gittes, F.; Schmidt, C. F. Biophys. J. 2003, 84, 1308−16. (26) Block, S. M. Optical Tweezers: A New Tool for Biophysics. In Noninvasive Techniques in Cell Biology; Wiley: New York, 1990. (27) Robblee, J. P.; Cao, W.; Henn, A.; Hannemann, D. E.; De La Cruz, E. M. Biochemistry 2005, 44, 10238−49. (28) Savin, A. V.; Mazo, M. A.; Kikot, I. P.; Manevitch, L. I.; Onufriev, A. V. Phys. Rev. B: Condens. Matter 2010, 83, 245406. (29) Takakura, Y.; Tsunashima, M.; Suzuki, J.; Usami, S.; Kakuta, Y.; Okino, N.; Ito, M.; Yamamoto, T. FEBS J. 2009, 276, 1383−97. (30) Phillips, R.; Kondev, J.; Theriot, J. Physical Biology of the Cell; Garland Science: New York, 2009. (31) Feynman, R. P. The Feynman Lectures on Physics; AddisonWesley: Reading, MA, 1963; Vol. 1. (32) Kerssemakers, J. W.; Munteanu, E. L.; Laan, L.; Noetzel, T. L.; Janson, M. E.; Dogterom, M. Nature 2006, 442, 709−12.
F
DOI: 10.1021/nl5049059 Nano Lett. XXXX, XXX, XXX−XXX
