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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.

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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

Kristensen, T. G., Nielsen, J. & Pedersen, C. N. S. (2010). A tree-based method for the rapid screening of chemical fingerprints. Algorithms for Molecular Biology, 5(9). https://doi.org/10.1186/1748-7188-5-9

Nielsen, K. H. & Andersen, C. (2010). AU-forskning på tværs. UNIvers, (6), 11.

Balog, R., Jørgensen, B., Nilsson, L., Andersen, M., Rienks, E., Bianchi, M., Fanetti, M., Lægsgaard, E., Baraldi, A., Lizzit, S., Sljivancanin, Z., Besenbacher, F., Hammer, B., Pedersen, T. G., Hofmann, P. & Hornekær, L. (2010). Bandgap opening in graphene induced by patterned hydrogen adsorption. Nature Materials, 9, 315-319. https://doi.org/10.1038/NMAT2710

Yde, J. C., Finster, K., Raiswell, R., Steffensen, J. P., Heinemeier, J., Olsen, J., Gunnlaugsson, H. P. & Nielsen, O. B. (2010). Basal ice microbiology at the margin of the Greenland ice sheet. Annals of Glaciology, 51(56), 71-79.

Nielsen, S. B., Franzmann, M., Basaiawmoit, R. V., Wimmer, R., Mikkelsen, J. D. & Otzen, D. E. (2010). beta-Sheet aggregation of kisspeptin-10 is stimulated by heparin but inhibited by amphiphiles. Biopolymers, 93(8), 678-89. https://doi.org/10.1002/bip.21434

Finster, K., Yde, J. C., Bárcena, T. G., Hodson, A. J., Mitchell, A. C., Knudsen, N. T., Hasholt, B., Nielsen, O. B., Gunnlaugsson, H. P. & Nørnberg, P. (2010). Biogeochemical studies of glacial ecosystems. Poster session presented at IPY symposium, København, Denmark.

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