Imaging microtubules and kinesin decorated microtubules using tapping mode atomic force microscopy in fluids |
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Authors: | Claudia M Kacher Ingrid M Weiss Russell J Stewart Christoph F Schmidt Paul K Hansma Manfred Radmacher Monika Fritz |
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Institution: | (1) Physik Department der TU-München, Institut für Biophysik E22, James-Franck-Strasse, D-85747 Garching, Germany e-mail: mfritz@physik.tu-muenchen.de, DE;(2) Bioengineering Department, University of Utah, Salt Lake City, UT 84112, USA, US;(3) Department of Biophysics and Physics of Complex Systems, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands, NL;(4) Department of Physics, University of California Santa Barbara, CA 93106, USA, US;(5) Sektion Physik der LMU München, Institut für Angewandte Physik, Amalienstrasse 54, D-80799 Munich, Germany, DE |
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Abstract: | 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.
Received: 23 February 1999 / Revised version: 19 July 1999 / Accepted: 17 August 1999 |
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Keywords: | Kinesin Microtubules Atomic force microscopy |
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