Heavy Meromyosin Molecules Extending More Than 50 nm above Adsorbing Electronegative Surfaces
Artikel i vetenskaplig tidskrift, 2010

In the in vitro motility assay, actin filaments are propelled by surface-adsorbed myosin motors, or rather, myosin motor fragments such as heavy meromyosin (HMM). Recently, efforts have been made to develop actomyosin powered nanodevices on the basis of this assay but such developments are hampered by limited understanding of the HMM adsorption geometry. Therefore, we here investigate the HMM adsorption geometries on trimethylchlorosilane-[TMCS-] derivatized hydrophobic surfaces and on hydrophilic negatively charged surfaces (SiO2). The TMCS surface is of great relevance in fundamental studies of actomyosin and both surface substrates are important for the development of motor powered nanodevices. Whereas both the TMCS and SiO2 surfaces were nearly saturated with HMM (incubation at 120 mu g mL(-1)) there was little actin binding on SiO2 in the absence of ATP and no filament sliding in the presence of ATP. This contrasts with excellent actin-binding and motility on TMCS. Quartz crystal microbalance with dissipation (QCM-D) studies demonstrate a HMM layer with substantial protein mass up to 40 nm above the TMCS surface, considerably more than observed for myosin subfragment 1 (SI; 6 nm). Together with the excellent actin transportation on TMCS, this strongly suggests that HMM adsorbs to TMCS mainly via its most C-terminal tail part. Consistent with this idea, fluorescence interference contrast (FLIC) microscopy showed that actin filaments are held by HMM 38 +/- 2 nm above the TMCS-surface with the catalytic site, on averge, 20-30 nm above the surface. Viewed in a context with FLIC, QCM-D and TIRF results, the lack of actin motility and the limited actin binding on SiO2 shows that HMM adsorbs largely via the actin-binding region on this surface with the C-terminal coiled-coil tails extending > 50 nm into solution. The results and new insights from this study are of value, not only for the development of motor powered nanodevices but also for the interpretation of fundamental biophysical studies of actomyosin function and for the understanding of surface protein interactions in general.

motor function


interference-contrast microscopy

myosin subfragment-2




oxidized silicon

quartz-crystal microbalance

driven motility


M. Persson

Linnéuniversitetet, Kalmar

N. Albet-Torres

Linnéuniversitetet, Kalmar

L. Ionov


M. Sundberg

Linnéuniversitetet, Kalmar

Fredrik Höök

Chalmers, Teknisk fysik, Biologisk fysik

S. Diez


A. Mansson

Linnéuniversitetet, Kalmar

M. Balaz

Lunds universitet

Linnéuniversitetet, Kalmar


07437463 (ISSN) 15205827 (eISSN)

Vol. 26 12 9927-9936





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