| 1. |
- Bunk, Richard, et al.
(författare)
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Actomyosin motility on nanostructured surfaces
- 2003
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Ingår i: Biochemical and Biophysical Research Communications. - 1090-2104. ; 301:3, s. 783-788
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Tidskriftsartikel (refereegranskat)abstract
- We have here, for the first time, used nanofabrication techniques to reproduce aspects of the ordered actomyosin arrangement in a muscle cell. The adsorption of functional heavy meromyosin (HMM) to five different resist polymers was first assessed. One group of resists (MRL-6000.1XP and ZEP-520) consistently exhibited high quality motility of actin filaments after incubation with HMM. A second group (PMMA-200, PMMA-950, and MRI-9030) generally gave low quality of motility with only few smoothly moving filaments. Based on these findings electron beam lithography was applied to a bi-layer resist system with PMMA-950 on top of MRL-6000.1XP. Grooves (100-200 nm wide) in the PMMA layer were created to expose the MRL-6000.1XP surface for adsorption of HMM and guidance of actin filament motility. This guidance was quite efficient allowing no U-turns of the filaments and approximately 20 times higher density of moving filaments in the grooves than on the surrounding PMMA.
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| 2. |
- Bunk, Richard, et al.
(författare)
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Towards a 'nano-traffic' system powered by molecular motors
- 2003
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Ingår i: Microelectronic Engineering. - 1873-5568. ; 67-8, s. 899-904
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Tidskriftsartikel (refereegranskat)abstract
- In this work, we reconstructed in vitro the behavior of two motor proteins-myosin and actin-responsible for the mechanical action of muscle cells. By transferring this in vivo system to an artificial environment, we were able to study the interaction between the proteins in more detail, as well as investigating the central mechanism of force production. Nm-patterning by e-beam lithography (EBL) could restore parts of the in vivo protein order, essential for potential nanotechnological applications. Much work was put into establishing the necessary compatibility between the biological and nano-lithographical processes. A range of EBL-resists were tested for protein compatibility. One particular kind (MRL-6000.1XP) supported good actin filament motility, while another (PMMA-950) behaved in the opposite way. Taking advantage of these findings, nm-sized lines were created in a double-layer structure of the two resists. The lines were found to act as binding sites for myosin, and as rectifying guides for the linearized motion of actin filaments. Velocities around 5 mum/s were measured. (C) 2003 Elsevier Science B.V. All rights reserved.
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| 3. |
- Sundberg, M, et al.
(författare)
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Silanized surfaces for in vitro studies of actomyosin function and nanotechnology applications
- 2003
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Ingår i: Analytical Biochemistry. - : Elsevier BV. - 1096-0309 .- 0003-2697. ; 323:1, s. 127-138
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Tidskriftsartikel (refereegranskat)abstract
- We have previously shown that selective heavy meromyosin (HMM) adsorption to predefined regions of nanostructured polymer resist surfaces may be used to produce a nanostructured in vitro motility assay. However, actomyosin function was of lower quality than on conventional nitrocellulose films. We have therefore studied actomyosin function on differently derivatized glass surfaces with the aim to find a substitute for the polymer resists. We have found that surfaces derivatized with trimethylchlorosilane (TMCS) were superior to all other surfaces tested, including nitrocellulose. High-quality actin filament motility was observed up to 6 days after incubation with HMM and the fraction of motile actin filaments and the velocity of smooth sliding were generally higher on TMCS than on nitrocellulose. The actomyosin function on TMCS-derivatized glass and nitrocellulose is considered in relation to roughness and hydrophobicity of these surfaces. The results suggest that TMCS is an ideal substitute for polymer resists in the nanostructured in vitro motility assay. Furthermore, TMCS derivatized glass also seems to offer several advantages over nitrocellulose for HMM adsorption in the ordinary in vitro motility assay. (C) 2003 Elsevier Inc. All rights reserved.
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