DNA-lipid conjugates have beenused to tether lipid bilayer patches and giant unilamellar vesicles (GUVs)either to fixed anti-sense DNA sequences covalently attached to a support or tothe anti-sense DNA sequence displayed on a fluid supported bilayer [1].  Inthe first case, the tethers are fixed, while in the latter they can diffuse inthe region between the two bilayer surfaces.  In both cases, GUVs areinitially tethered, then, depending on the number density of DNAs on thesurface, the GUVs rupture to form patches.  The first system producesstable patches or stably attached GUVs independent of lipid composition, whilethe second system requires specific lipid compositions to achieve stability.?  Both systems can be used as substratesfor further manipulation using DNA-lipid fusion machinerywe have developed [2].  Thismachinery can also be used to deliver integral membrane proteins to GUVs orpatches.  Because the patches or GUVs are held away from the underlying support at a distance determined by the DNA length, unfavorable interactions with the support are avoided, especially important when working with integral membrane proteins.  When GUVs or patches aretethered to supported bilayers with mobile tethers of different lengths,interesting topological domains are formed where tethers of different lengthsegregate.  Advantages anddisadvantages of this approach compared with tethered vesicles [3] will be discussed. [1] M. Chung, R. D. Lowe, Y.-H.M. Chan, P. V. Ganesan and S. G. Boxer, J. Structural Biology, 168,190-199 (2009).[2]Y.-H. M. Chan, B. van Lengerich and S. G. Boxer, Biointerphases, 3,FA17-FA21 (2008); Y.-H. M.  Chan,B. van Lengerich, S. G.  Boxer, PNAS, 106, 979-984 (2009).[3] Y.-H. M. Chan, P. Lenz and S. G. Boxer, PNAS, 104, 18913-18918 (2007).

Tethered membrane patches and GUVs as tools in membrane biophysics

241st National Meeting (2011)

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