Journal
BIOPHYSICAL JOURNAL
Volume 105, Issue 10, Pages 2355-2365Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2013.10.007
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Funding
- Engineering and Physical Sciences Research Council [EP/I000623/1, EP/J01756611]
- EPSRC [EP/I000623/1, EP/J017566/1, EP/K023845/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/J017566/1, GR/S87195/01, EP/I000623/1, EP/K023845/1] Funding Source: researchfish
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We report on the use of supported lipid bilayers to reveal dynamics of actin polymerization from a nonpolymerizing subphase via cationic phospholipids. Using varying fractions of charged lipid, lipid mobility, and buffer conditions, we show that dynamics at the nanoscale can be used to control the self-assembly of these structures. In the case of fluid-phase lipid bilayers, the actin adsorbs to form a uniform two-dimensional layer with complete surface coverage whereas gel-phase bilayers induce a network of randomly oriented actin filaments, of lower coverage. Reducing the pH increased the polymerization rate, the number of nucleation events, and the total coverage of actin. A model of the adsorption/diffusion process is developed to provide a description of the experimental data and shows that, in the case of fluid-phase bilayers, polymerization arises equally due to the adsorption and diffusion of surface-bound monomers and the addition of monomers directly from the solution phase. In contrast, in the case of gel-phase bilayers, polymerization is dominated by the addition of monomers from solution. In both cases, the filaments are stable for long times even when the G-actin is removed from the supernatant making this a practical approach for creating stable lipid-actin systems via self-assembly.
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