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pubmed:abstractText |
Poly(methylmethacrylate) (PMMA), a photoresist polymer, was found to be useful for immobilizing heavy meromyosin (HMM) molecules while retaining their abilities to support the movement of actin filaments. PMMA substrate was spin-coated on a coverslip, and various shapes of PMMA tracks, such as straight lines, concentric circles, and alphabetical letters, were fabricated by UV photolithography. An observation by a Tapping mode atomic force microscope (AFM) shows that the typical circular tracks were 1-2 microns wide and about 200 nm high. In in vitro motility assay, a solution of HMM molecules was applied to immobilize the molecules on the tracks by adsorption, and movement of actin filaments labeled with tetramethylrhodamine-phalloidin were observed in the presence of ATP by using an epifluorescence microscope and an image-intensified CCD camera. Actin filaments were seen to move precisely only on the PMMA tracks, and their traces drew the exact shapes of the tracks. The mean velocity of actin movement on the PMMA was 4.5 mm/s at 25 degrees C, and it was comparable to that on a conventionally used nitrocellulose film.
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