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Distinct Network Morphologies from In Situ Polymerization of Microtubules in Giant Polymer-Lipid Hybrid Vesicles

Creating artificial cells with a dynamic cytoskeleton, akin to those in living cells, is a major goal in bottom-up synthetic biology. In this study, we demonstrate the in situ polymerization of microtubules encapsulated in giant polymer-lipid hybrid vesicles (GHVs) composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine and an amphiphilic block copolymer. The block copolymer is comprised of poly(cholesteryl methacrylate-co-butyl methacrylate) as the hydrophobic block and either poly(6-O-methacryloyl-D-galactopyranose) or poly(carboxyethyl acrylate) as the hydrophilic extension. Depending on the concentrations of guanosine triphosphate (GTP) or its slowly hydrolyzable analog, guanosine-5′-[(α,β)-methyleno]triphosphate (GMPCPP), different microtubule morphologies are observed, including encapsulated microtubule networks, spike protrusions, as well as membrane-associated or aggregated microtubules. Overall, this work represents a step forward in mimicking the cellular cytoskeletons and uncovering the influence of membrane composition on microtubule morphologies.

Recent publications by network

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Otzen, D. E. (2010). Amyloid formation in surfactants and alcohols: membrane mimetics or structural switchers? Current protein & peptide science, 11(5), 355-71.

Pedersen, J. S., Andersen, C. B. & Otzen, D. (2010). Amyloid structure--one but not the same: the many levels of fibrillar polymorphism. FEBS journal, 277(22), 4591-4601. https://doi.org/10.1111/j.1742-4658.2010.07888.x

Feliu, E., Knudsen, M., Wiuf, C. H. & Andersen, L. N. (2010). An Algebraic Approach to Signaling Cascades with n Layers. Poster session presented at The 11th International Conference on Systems Biology, Edinburgh, United Kingdom.

Giehm, L., Dal Degan, F., Fraser, P., Klysner, S. & Otzen, D. E. (2010). An Aß concatemer with altered aggregation propensities. BBA General Subjects, 1804(10), 2025-35. https://doi.org/10.1016/j.bbapap.2010.06.023

Michelow, I. C., Dong, M., Mungall, B. A., Yantosca, L. M., Lear, C., Ji, X., Karpel, M., Rootes, C. L., Brudner, M., Houen, G., Eisen, D. P., Kinane, T. B., Takahashi, K., Stahl, G. L., Olinger, G. G., Spear, G. T., Ezekowitz, R. A. B. & Schmidt, E. V. (2010). A novel L-ficolin/mannose-binding lectin chimeric molecule with enhanced activity against Ebola virus. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.M110.106260

Subramani, R., Jensen, S. J., Rotaru, A., Andersen, F. F., Gothelf, K. V., Mamdouh, W., Besenbacher, F., Dong, M. & Knudsen, B. R. (2010). A Novel Secondary DNA Binding Site in Human Topoisomerase I Unravelled by using a 2D DNA Origami Platform. A C S Nano, 4, 5969-77.

Schaefer, A., Im, H.-I., Venø, M. T., Fowler, C. D., Min, A., Intrator, A., Kjems, J., Kenny, P. J., O'Carroll, D. & Greengard, P. (2010). Argonaute 2 in dopamine 2 receptor-expressing neurons regulates cocaine addiction. Journal of Experimental Medicine, 207(9), 1843-51. https://doi.org/10.1084/jem.20100451

Bramsen, J. B., Pakula, M. M., Hansen, T. B., Bus, C., Langkjær, N., Odadzic, D., Smicius, R., Wengel, S. L., Chattopadhyaya, J., Engels, J. W., Herdewijn, P., Wengel, J. & Kjems, J. (2010). A screen of chemical modifications identifies position-specific modification by UNA to most potently reduce siRNA off-target effects. Nucleic Acids Research, 38(17), 5761-5773. https://doi.org/10.1093/nar/gkq341

Giehm, L., Lorenzen, N. & Otzen, D. E. (2010). Assays for α-synuclein aggregation. Methods. https://doi.org/10.1016/j.ymeth.2010.12.008

Jeppesen, M. D., Hein, K., Nissen, P., Westh, P. & Otzen, D. E. (2010). A thermodynamic analysis of fibrillar polymorphism. Advances in Biophysical Chemistry, 149(1-2), 40-6. https://doi.org/10.1016/j.bpc.2010.03.016

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