Imaging microtubules and kinesin decorated microtubules using tapping mode atomic force microscopy in fluids.

Published

Journal Article

The atomic force microscope has been used to investigate microtubules and kinesin decorated microtubules in aqueous solution adsorbed onto a solid substrate. The netto negatively charged microtubules did not adsorb to negatively charged solid surfaces but to glass covalently coated with the highly positively charged silane trimethoxysilylpropyldiethylenetriamine (DETA) or a lipid bilayer of 1,2-dipalmitoyl-3-dimethylammoniumpropane. Using electron beam deposited tips for microtubules adsorbed on DETA, single protofilaments could be observed showing that the resolution is up to 5 nm. Under conditions where the silane coated surfaces are hydrophobic, microtubules opened, presumably at the seam, whose stability is lower than that of the bonds between the other protofilaments. This led to a "sheet" with a width of about 100 nm firmly attached to the surface. Microtubules decorated with a stoichiometric low amount of kinesin molecules in the presence of the non-hydrolyzable ATP-analog 5'-adenylylimidodiphosphate could also be adsorbed onto silane-coated glass. Imaging was very stable and the molecules did not show any scan-induced deformation even after hundreds of scans with a scan frequency of 100 Hz.

Full Text

Duke Authors

Cited Authors

  • Kacher, CM; Weiss, IM; Stewart, RJ; Schmidt, CF; Hansma, PK; Radmacher, M; Fritz, M

Published Date

  • January 2000

Published In

Volume / Issue

  • 28 / 8

Start / End Page

  • 611 - 620

PubMed ID

  • 10663528

Pubmed Central ID

  • 10663528

Electronic International Standard Serial Number (EISSN)

  • 1432-1017

International Standard Serial Number (ISSN)

  • 0175-7571

Digital Object Identifier (DOI)

  • 10.1007/s002490050001

Language

  • eng