Supported Membranes

Supported Lipid Bilayers as Model Systems for Lipid-Protein Interactions

We have a long-standing interest in the further development of supported lipid bilayers as models of biological membranes. Having pioneered supported membranes in the 1980s, we developed methods to prepare single planar lipid bilayers on soft polymer cushions. Several integral membrane proteins have been shown to be laterally mobile in these model membranes. Their motions and interactions can be studied by single-molecule total internal reflection fluorescence microscopy (TIRFM), by fluorescence recovery after photobleaching (FRAP), and by fluorescence interference contrast (FLIC) microscopy.

This system is also very useful to study the assembly and dynamics of lateral membrane domains, “lipid rafts” . For example, we recently discovered that, in asymmetric bilayers, raft-like lipid domains are induced in inner leaflet lipid mixtures by raft-like lipid domains in outer leaflet mixtures in asymmetric bilayers. We currently study the targeting of proteins described in our other research areas to different domains in these heterogeneous membrane models.

: Design of a tethered polymer-supported lipid bilayer.

: Coupled asymmetric lipid membrane domains with 20% cholesterol in both leaflets.

Recent Publications:

Kreutzberger AJB, Kiessling V, Liang B, Yang ST, Castle JD, Tamm LK. Asymmetric Phosphatidylethanolamine Distribution Controls Fusion Pore Lifetime and Probability. Biophys J. 2017 Oct 13. pii: S0006-3495(17)31026-3. doi: 10.1016/j.bpj.2017.09.014. [Epub ahead of print](https://www.ncbi.nlm.nih.gov/pubmed/29037600)

Kiessling V, Liang B, Kreutzberger AJ, Tamm LK. Planar Supported Membranes with Mobile SNARE Proteins and Quantitative Fluorescence Microscopy Assays to Study Synaptic Vesicle Fusion. Front Mol Neurosci. 2017 Mar 16;10:72. doi: 10.3389/fnmol.2017.00072. eCollection 2017. (https://www.ncbi.nlm.nih.gov/pubmed/28360838)

Yang ST, Lim S, Kiessling V, Kwon I, Tamm LK. (2016) Site-specific fluorescent labeling to visualize membrane translocation of a myristoyl switch protein. Sci Rep. 2016 Sep 8;6:32866. doi: 10.1038/srep32866.( https://www.ncbi.nlm.nih.gov/pubmed/27605302)

Kiessling V, Yang ST, Tamm LK. Supported lipid bilayers as models for studying membrane domains. Curr Top Membr. 2015 April;75:1-23 (http://www.ncbi.nlm.nih.gov/pubmed/26015279)

Moissoglu, K., Kiessling, V., Wan, C., Hoffman, B.D., Norambuena, A., Tamm, L.K., Schwartz, M.A. Regulation of Rac Translocation and Activation by Membrane Domains and Their Boundaries. Journal of Cell Science, 2014, Apr 2. [Epub ahead of print] (http://www.ncbi.nlm.nih.gov/pubmed/24695858)

Wan, C., Kiessling, V., Cafiso, D.S., and Tamm, L.K. (2011) Partitioning of synaptotagmin I C2 domains between liquid-ordered and liquid-disordered inner leaflet lipid phases. Biochemistry 50:2478-2485. (http://www.ncbi.nlm.nih.gov/pubmed/21322640)

Kiessling, V., Domanska, M.K., and Tamm, L.K. (2010) Single SNARE-mediated vesicle fusion observedin vitro by polarized TIRFM. Biophys. J. 99:4047-4055. (http://www.ncbi.nlm.nih.gov/pubmed/21156148)

Murray, D.H., Tamm, L.K., Kiessling, V.(2009), Supported double membranes. J. Structural Biology 168, 183-189 (http://www.ncbi.nlm.nih.gov/pubmed/19236921)

Kiessling, V., Wan, C., and Tamm, L.K. (2009). Domain coupling in asymmetric lipid bilayers. Biochimica et Biophysica Acta, 1788: 64-71. (http://www.ncbi.nlm.nih.gov/pubmed/18848518)

Wan, C., Kiessling, V., and Tamm, L.K.(2008). Coupling of cholesterol-rich lipid phases in asymmetric bilayers. Biochemistry 47: 2190-2198. (http://www.ncbi.nlm.nih.gov/pubmed/18215072)

Kiessling, V., Domanska, M.K., Murray, D., Wan, C., and Tamm L.K.(2008). Supported lipid bilayers: development and application in chemical biology. Wiley Encyclopedia of Chemical Biology, John Wiley & Sons, Hoboken, Volume 4, pp. 411-422.

Kiessling, V., Crane, J.M., Tamm L.K. (2006). Transbilayer effects of raft-like lipid domains in asymmetric planar bilayer measured by single molecule tracking. Biophysical Journal. 91: 3313-3326. (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1614489/)

Crane, J.M., V. Kiessling, and L.K. Tamm (2005). Measuring lipid asymmetry in planar supported bilayers by fluorescence interference contrast microscopy. Langmuir. 21: 1377-1388. (http://www.ncbi.nlm.nih.gov/pubmed/15697284)

Crane, J.M. and L.K. Tamm (2004). Role of cholesterol in the formation and nature of lipid rafts in planar and spherical model membranes. Biophys. J. 86: 2965-2979. (http://www.ncbi.nlm.nih.gov/pubmed/15111412)

Kiessling, V. and L.K. Tamm (2003). Measuring distances in supported bilayers by fluorescence interference-contrast microscopy: polymer supports and SNARE proteins. Biophys. J. 84: 408-418. (http://www.ncbi.nlm.nih.gov/pubmed/12524294)

Wagner, M.L. and L.K. Tamm (2001). Reconstituted syntaxin1A/SNAP25 interacts with negatively charged lipids as measured by lateral diffusion in planar supported bilayers. Biophys. J. 81: 266-275.(http://www.ncbi.nlm.nih.gov/pubmed/11423412)

Tamm, L.K., V. Kiessling, and M.L. Wagner (2001). Membrane dynamics. Encyclopedia of Life Sciences.Nature Publishing Group, Macmillan Publishers, Ltd. (http://www.els.net)

Wagner, M.L. and L.K. Tamm (2000). Tethered polymer-supported planar lipid bilayers for reconstitution of integral membrane proteins: silane-PEG-lipid as a cushion and covalent linker. Biophys. J.79:1400-1414. (http://www.ncbi.nlm.nih.gov/pubmed/10969002)