Kinesin Walks Hand-Over-Hand

Science

Volumn 303, Issue 5658, 2004, Pages 676-678

  Yildiz, Ahmet ^a   Tomishige, Michio ^c,d   Vale, Ronald D ^c   Selvin, Paul R ^a,b  

^a UNIVERSITY OF ILLINOIS AT URBANA CHAMPAIGN   (United States)

^b University of Illinois at Urbana Champaign   (United States)

^c UNIVERSITY OF CALIFORNIA   (United States)

^d UNIVERSITY OF TOKYO   (Japan)

Author keywords

[No Author keywords available]

Indexed keywords

DIMERS; HYDROLYSIS; PHOSPHATES;

KINESIN;

PROTEINS;

ADENOSINE TRIPHOSPHATE; DYE; KINESIN;

LOCOMOTION;

ARTICLE; HYDROLYSIS; KINETICS; MICROTUBULE; PRIORITY JOURNAL; PROTEIN BINDING;

ADENOSINE TRIPHOSPHATE; CARBOCYANINES; DIMERIZATION; FLUORESCENCE; FLUORESCENT DYES; HUMANS; KINESIN; KINETICS; MICROTUBULES; MODELS, BIOLOGICAL; MODELS, MOLECULAR; MOLECULAR MOTOR PROTEINS; MUTATION; PROTEIN CONFORMATION;

EID: 0942279641     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1093753     Document Type: Article

References (24)

1. J. Howard, A. J. Hudspeth, R. D. Vale, Nature 342, 154 (1989).
2. K. Svoboda, C. F. Schmidt, B. J. Schnapp, S. M. Block, Nature 365, 721 (1993).
3. R. D. Vale, R. A. Milligan, Science 288, 88 (2000).
4. L. S. Goldstein, A. V. Philp, Annu. Rev. Cell Dev. Biol. 15, 141 (1999).
5. E. Reid et al., Am. J. Hum. Genet. 71, 1189 (2002).
6. F. Kozielski et al., Cell 91, 985 (1997).
7. S. Rice et al., Nature 402, 778 (1999).
8. R. D. Vale, R. J. Fletterick, Annu. Rev. Cell Dev. Biol. 13, 745 (1997).
9. W. Hua, J. Chung, J. Gelles, Science 295, 844 (2002).
10. A. Hoenger et al., J. Mol. Biol. 297, 1087 (2000).
11. A. Yildiz et al., Science 300, 2061 (2003).
12. For unpolarized dyes, the center of the PSF corresponds to the dye position. For dyes with fixed orientation that emit polarized fluorescence, the PSF can take unusual shapes, particularly when viewed with spherical aberrations, and the PSF center does not necessarily correspond to the dye position (23). In our case, the PSF is well fit to a symmetric two-dimensional Gaussian, implying the center of the PSF corresponds to the dye position. Furthermore, the orientation of the dye in both the hand-over-hand and inchworm models is expected to be unchanged when on the front and rear head, and hence there should be no effect of PSF shape or dye orientation on step-size determination.
13. Materials and methods are available as supporting material on Science Online.
14. The uncertainty, or the standard error of the mean, of the PSF centers for relatively bright spots (5000 photons per image) is <3 nm. Bright spots (>7000 photons per image) yield ±2 nm localization of PSF centers (from seven molecules; traces are shown in Fig. 2). We determined the step size by calculating the distance between average positions of plateaus before and after a step. The standard error of the mean of two adjacent plateaus gives the precision of the step size (11).
15. K. Kawaguchi, S. Ishiwata, Science 291, 667 (2001).
16. K. Kawaguchi, S. Uemura, S. Ishiwata, Biophys. J. 84, 1103 (2003).
17. S. S. Rosenfeld, P. M. Fordyce, G. M. Jefferson, P. H. King, S. M. Block, J. Biol. Chem. 278, 18550 (2003).
18. A. B. Asenjo, N. Krohn, H. Sosa, Nature Struct. Biol. 10, 836 (2003).
19. J. N. Forkey, M. E. Quinlan, M. Alexander Shaw, J. E. T. Corrie, Y. E. Goldman, Nature 422, 399 (2003).
20. M. Y. Ali et al., Nature Struct. Biol. 9, 464 (2002).
21. S. M. Block, C. L. Asbury, J. W. Shaevitz, M. J. Lang, Proc. Natl. Acad. Sci. U.S.A. 100, 2351 (2003).
22. R. Koradi, M. Billeter, K. Wüthrich, J. Mol. Graph. 14, 51 (1996).
23. A. P. Bartko, R. M. Dickson, J. Phys. Chem. B 103 11237 (1999).
24. We thank Y. Oono for helpful discussion of one. versus two-headed binding and the use of fig. S2 and A. Carter for preparing Fig. 1B. Supported by NIH grant nos. AR44420 (P.R.S.) and AR42895 (R.D.V.).