Rapid And Efficient Co-reconstitution Of Bacterial Mechanosensitive Ion Channels Of Small And Large Conductance Into Liposomes

Abstract

Bacterial mechanosensitive (MS) channels protect bacterial cells from osmotic shock, acting as emergency relief valves (1,2). E. coli has three such channels, the MS channel of large conductance (MscL), the MS channel of small conductance (MscS) and the MS channel of mini conductance (MscM). Both MscL and MscS have been extensively studied using the patch-clamp technique in giant spheroplasts (3,4). However, only MscL incorporates efficiently in liposomes (2,5). Here we report the first example of co-reconstitution of both MscS and MscL into azolectin liposomes. We also report reconstitution efficiencies of both proteins into liposomes of different lipid composition and using different incorporation methods.View Large Image | View Hi-Res Image | Download PowerPoint Slide1. Martinac, B., J. Cell Sci., 117, 2449-2460 (2004).2. Perozo, E., Nat. Rev. Mol. Biol., 7, 109-119 (2006).3. Martinac, B., Buechner, M., Delcour, A.H., Adler, J., Kung, C. PNAS, 84, 2297-2301 (1987).4. Booth, I.R., Edwards, M.D., Black, S., Schumann, U., Miller, S. Nat. Rev. Microbiol., 5, 431-440 (2007).5. Hase, C.C., Le Dain, A.C., Martinac, B., J Biol. Chem, 270, 18329-18334 (1995).Supported by the Australian Research Council and National Health & Medical Research Council.