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Selective spatial localization of actomyosin motor function by chemical surface patterning

Sundberg, Mark ; Balaz, Martina ; Bunk, Richard LU ; Rosengren-Holmberg, Jenny P. ; Montelius, Lars LU ; Nicholls, Ian A. ; Omling, Pär LU ; Tagerud, Sven and Mansson, Alf (2006) In Langmuir 22(17). p.7302-7312
Abstract
We have previously described the efficient guidance and unidirectional sliding of actin filaments along nanosized tracks with adsorbed heavy meromyosin (HMM; myosin II motor fragment). In those experiments, the tracks were functionalized with trimethylchlorosilane (TMCS) by chemical vapor deposition (CVD) and surrounded by hydrophilic areas. Here we first show, using in vitro motility assays on nonpatterned and micropatterned surfaces, that the quality of HMM function on CVD-TMCS is equivalent to that on standard nitrocellulose substrates. We further examine the influences of physical properties of different surfaces (glass, SiO2, and TMCS) and chemical properties of the buffer solution on motility. With the presence of methylcellulose in... (More)
We have previously described the efficient guidance and unidirectional sliding of actin filaments along nanosized tracks with adsorbed heavy meromyosin (HMM; myosin II motor fragment). In those experiments, the tracks were functionalized with trimethylchlorosilane (TMCS) by chemical vapor deposition (CVD) and surrounded by hydrophilic areas. Here we first show, using in vitro motility assays on nonpatterned and micropatterned surfaces, that the quality of HMM function on CVD-TMCS is equivalent to that on standard nitrocellulose substrates. We further examine the influences of physical properties of different surfaces (glass, SiO2, and TMCS) and chemical properties of the buffer solution on motility. With the presence of methylcellulose in the assay solution, there was HMM-induced actin filament sliding on both glass/SiO2 and on TMCS, but the velocity was higher on TMCS. This difference in velocity increased with decreasing contact angles of the glass and SiO2 surfaces in the range of 20-67 degrees (advancing contact angles for water droplets). The corresponding contact angle of CVD-TMCS was 81 degrees. In the absence of methylcellulose, there was high-quality motility on TMCS but no motility on glass/SiO2. This observation was independent of the contact angle of the glass/SiO2 surfaces and of HMM incubation concentrations (30-150 mu g mL(-1)) and ionic strengths of the assay solution (20-50 mM). Complete motility selectivity between TMCS and SiO2 was observed for both nonpatterned and for micro- and nanopatterned surfaces. Spectrophotometric analysis of HMM depletion during incubation, K/EDTA ATPase measurements, and total internal reflection fluorescence spectroscopy of HMM binding showed only minor differences in HMM surface densities between TMCS and SiO2/glass. Thus, the motility contrast between the two surface chemistries seems to be attributable to different modes of HMM binding with the hindrance of actin binding on SiO2/glass. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Langmuir
volume
22
issue
17
pages
7302 - 7312
publisher
The American Chemical Society (ACS)
external identifiers
  • wos:000239596300036
  • scopus:33748561805
  • pmid:16893230
ISSN
0743-7463
DOI
10.1021/la060365i
language
English
LU publication?
yes
id
6c6fc436-7e43-4891-bc76-a2ea70370008 (old id 398356)
date added to LUP
2016-04-01 12:30:41
date last changed
2021-09-01 02:25:06
@article{6c6fc436-7e43-4891-bc76-a2ea70370008,
  abstract     = {We have previously described the efficient guidance and unidirectional sliding of actin filaments along nanosized tracks with adsorbed heavy meromyosin (HMM; myosin II motor fragment). In those experiments, the tracks were functionalized with trimethylchlorosilane (TMCS) by chemical vapor deposition (CVD) and surrounded by hydrophilic areas. Here we first show, using in vitro motility assays on nonpatterned and micropatterned surfaces, that the quality of HMM function on CVD-TMCS is equivalent to that on standard nitrocellulose substrates. We further examine the influences of physical properties of different surfaces (glass, SiO2, and TMCS) and chemical properties of the buffer solution on motility. With the presence of methylcellulose in the assay solution, there was HMM-induced actin filament sliding on both glass/SiO2 and on TMCS, but the velocity was higher on TMCS. This difference in velocity increased with decreasing contact angles of the glass and SiO2 surfaces in the range of 20-67 degrees (advancing contact angles for water droplets). The corresponding contact angle of CVD-TMCS was 81 degrees. In the absence of methylcellulose, there was high-quality motility on TMCS but no motility on glass/SiO2. This observation was independent of the contact angle of the glass/SiO2 surfaces and of HMM incubation concentrations (30-150 mu g mL(-1)) and ionic strengths of the assay solution (20-50 mM). Complete motility selectivity between TMCS and SiO2 was observed for both nonpatterned and for micro- and nanopatterned surfaces. Spectrophotometric analysis of HMM depletion during incubation, K/EDTA ATPase measurements, and total internal reflection fluorescence spectroscopy of HMM binding showed only minor differences in HMM surface densities between TMCS and SiO2/glass. Thus, the motility contrast between the two surface chemistries seems to be attributable to different modes of HMM binding with the hindrance of actin binding on SiO2/glass.},
  author       = {Sundberg, Mark and Balaz, Martina and Bunk, Richard and Rosengren-Holmberg, Jenny P. and Montelius, Lars and Nicholls, Ian A. and Omling, Pär and Tagerud, Sven and Mansson, Alf},
  issn         = {0743-7463},
  language     = {eng},
  number       = {17},
  pages        = {7302--7312},
  publisher    = {The American Chemical Society (ACS)},
  series       = {Langmuir},
  title        = {Selective spatial localization of actomyosin motor function by chemical surface patterning},
  url          = {http://dx.doi.org/10.1021/la060365i},
  doi          = {10.1021/la060365i},
  volume       = {22},
  year         = {2006},
}