Photoactivatable Lipids

  • Photoactivatable lipids: A key tool to find proteins that interact specifically with lipids.

Interactions between proteins and lipids find increasing interest as a mean of regulating cellular functions. Until recently their study in living cells were nearly impossible. Therefore, we developed photoactivatable radioactive lipids that allow discovery of the protein interaction partners. These lipids contain a diazirine ring that can be activated by UV-light to form covalent links to any interaction partner (Thiele et al. 2000).

Since the photoactivatable lipids are radioactively labeled, interactions are easily detected by a SDS-PAGE/autoradiography.

After we originally described the method  [I.] we and many others have successfully used the technology to demonstrate specific interaction between proteins and certain lipids . [II.-XI.]

I. Thiele, C., M.J. Hannah, F. Fahrenholz, and W.B. Huttner, Cholesterol binds to synaptophysin and is required for biogenesis of synaptic vesicles. Nat Cell Biol, 2000. 2(1): p. 42-9.

II. Lecat, S., P. Verkade, C. Thiele, K. Fiedler, K. Simons, and F. Lafont, Different properties of two isoforms of annexin XIII in MDCK cells. J Cell Sci, 2000.113(Pt 14): p. 2607-18.

III. Simons, M., E.M. Kramer, C. Thiele, W. Stoffel, and J. Trotter, Assembly of myelin by association of proteolipid protein with cholesterol- and galactosylceramide-rich membrane domains. J Cell Biol, 2000. 151(1): p. 143-54.

IV. Matyash, V., et al., Distribution and transport of cholesterol in Caenorhabditis elegans. Mol Biol Cell, 2001. 12(6): p. 1725-36.

V. Lang, T., et al., SNAREs are concentrated in cholesterol-dependent clusters that define docking and fusion sites for exocytosis. Embo J, 2001. 20(9): p. 2202-13.

VI. Wang, N., D.L. Silver, C. Thiele, and A.R. Tall, ATP-binding cassette transporter A1 (ABCA1) functions as a cholesterol efflux regulatory protein. J Biol Chem, 2001. 276(26): p. 23742-7.

VII. Smit, J.M., B.L. Waarts, K. Kimata, W.B. Klimstra, R. Bittman, and J. Wilschut, Adaptation of alphaviruses to heparan sulfate: interaction of Sindbis and Semliki forest viruses with liposomes containing lipid-conjugated heparin. J Virol, 2002. 76(20): p. 10128-37.

VIII. Mintzer, E.A., B.L. Waarts, J. Wilschut, and R. Bittman, Behavior of a photoactivatable analog of cholesterol, 6-photocholesterol, in model membranes.FEBS Lett, 2002. 510(3): p. 181-4.

IX. Charrin, S., et al., A physical and functional link between cholesterol and tetraspanins. Eur J Immunol, 2003. 33(9): p. 2479-89.

X. Eroglu, C., B. Brugger, F. Wieland, and I. Sinning, Glutamate-binding affinity of Drosophila metabotropic glutamate receptor is modulated by association with lipid rafts. Proc Natl Acad Sci U S A, 2003. 100(18): p. 10219-24.

XI. Adams, C.M., et al., Cholesterol and 25-hydroxycholesterol inhibit activation of SREBPs by different mechanisms, both involving SCAP and insigs. JBiol Chem, 2004.

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