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High resolution two-dimensional detection of ncd motility with optical tweezers

Publication ,  Conference
Allersma, MW; Gittes, F; deCastro, MJ; Stewart, RJ; Schmidt, CF
Published in: Materials Research Society Symposium - Proceedings
December 1, 1998

The ATP-dependent motility of the kinesin-related non claret disjunctional (ncd) mechanoenzyme was observed in an in vitro bead motility assay using optical tweezers in combination with a new two-dimensional displacement detection method. The detection technique is based on observing the far-field interference pattern formed in the back focal plane (BFP) of the microscope condenser by the illuminating laser focus and the light scattered from the trapped dielectric bead. The ability to observe the two-dimensional motion, with high temporal and spatial resolution, and in a manner largely independent of position in the microscope field-of-view, is the particular advantage of this detection method. In the assay, a fusion protein (GST-N195) of truncated ncd and glutathione-S-transferase was adsorbed to silica beads and the axial and lateral motions of the beads along the microtubule surface were observed. The average axial velocity of the ncd coated beads was 230±30 nm/s (±std. dev.). Spectral analysis of bead motion showed an increase in viscous drag near the surface. Furthermore, we also found that any elastic constraints of the moving motors are much smaller than the constraints due to binding in the presence of the non-hydrolyzable nucleotide adenylylimido-diphosphate (AMP-PNP).

Duke Scholars

Published In

Materials Research Society Symposium - Proceedings

ISSN

0272-9172

Publication Date

December 1, 1998

Volume

489

Start / End Page

85 / 90
 

Citation

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Allersma, M. W., Gittes, F., deCastro, M. J., Stewart, R. J., & Schmidt, C. F. (1998). High resolution two-dimensional detection of ncd motility with optical tweezers. In Materials Research Society Symposium - Proceedings (Vol. 489, pp. 85–90).
Allersma, M. W., F. Gittes, M. J. deCastro, R. J. Stewart, and C. F. Schmidt. “High resolution two-dimensional detection of ncd motility with optical tweezers.” In Materials Research Society Symposium - Proceedings, 489:85–90, 1998.
Allersma MW, Gittes F, deCastro MJ, Stewart RJ, Schmidt CF. High resolution two-dimensional detection of ncd motility with optical tweezers. In: Materials Research Society Symposium - Proceedings. 1998. p. 85–90.
Allersma, M. W., et al. “High resolution two-dimensional detection of ncd motility with optical tweezers.” Materials Research Society Symposium - Proceedings, vol. 489, 1998, pp. 85–90.
Allersma MW, Gittes F, deCastro MJ, Stewart RJ, Schmidt CF. High resolution two-dimensional detection of ncd motility with optical tweezers. Materials Research Society Symposium - Proceedings. 1998. p. 85–90.

Published In

Materials Research Society Symposium - Proceedings

ISSN

0272-9172

Publication Date

December 1, 1998

Volume

489

Start / End Page

85 / 90