Proceedings of the National Academy of Sciences - PNAS, 2011-04, Vol.108 (15), p.6264-6269
2011
Details
- Autor(en) / Beteiligte
- Titel
- Cardiolipin microdomains localize to negatively curved regions of Escherichia coli membranes
- Ist Teil von
- Proceedings of the National Academy of Sciences - PNAS, 2011-04, Vol.108 (15), p.6264-6269
- Ort / Verlag
- United States: National Academy of Sciences
- Erscheinungsjahr
- 2011
- Link zum Volltext
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Many proteins reside at the cell poles in rod-shaped bacteria. Several hypotheses have drawn a connection between protein localization and the large cell-wall curvature at the poles. One hypothesis has centered on the formation of microdomains of the lipid cardiolipin (CL), its localization to regions of high membrane curvature, and its interaction with membrane-associated proteins. A lack of experimental techniques has left this hypothesis unanswered. This paper describes a microtechnology-based technique for manipulating bacterial membrane curvature and quantitatively measuring its effect on the localization of CL and proteins in cells. We confined Escherichia coli spheroplasts in microchambers with defined shapes that were embossed into a layer of polymer and observed that the shape of the membrane deformed predictably to accommodate the walls of the microchambers. Combining this technique with epifluorescence microscopy and quantitative image analyses, we characterized the localization of CL microdomains in response to E. coli membrane curvature. CL microdomains localized to regions of high intrinsic negative curvature imposed by microchambers. We expressed a chimera of yellow fluorescent protein fused to the N-terminal region of MinD--a spatial determinant of E. coli division plane assembly--in spheroplasts and observed its colocalization with CL to regions of large, negative membrane curvature. Interestingly, the distribution of MinD was similar in spheroplasts derived from a CL synthase knockout strain. These studies demonstrate the curvature dependence of CL in membranes and test whether these structures participate in the localization of MinD to regions of negative curvature in cells.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0027-8424eISSN: 1091-6490DOI: 10.1073/pnas.1015757108Titel-ID: cdi_proquest_miscellaneous_907160927
- Format
- –
- Schlagworte
- Adenosine Triphosphatases - analysis, Adenosine Triphosphatases - genetics, Adenosine Triphosphatases - metabolism, Bacteria, Bacterial proteins, Bacterial Proteins - analysis, Bacterial Proteins - genetics, Bacterial Proteins - metabolism, Biochemistry, Biological Sciences, cardiolipin, Cardiolipins - chemistry, Cardiolipins - metabolism, Cell Division, Cell membranes, Chimeras, Curvature, E coli, Escherichia coli, Escherichia coli - chemistry, Escherichia coli - metabolism, Escherichia coli - ultrastructure, Escherichia coli Proteins - analysis, Escherichia coli Proteins - genetics, Escherichia coli Proteins - metabolism, fluorescence microscopy, Gene Knockout Techniques, Geodetic position, Image processing, Lipid rafts, Lipids, Luminescent Proteins - analysis, Luminescent Proteins - genetics, Luminescent Proteins - metabolism, Membrane Microdomains - chemistry, Membrane Microdomains - metabolism, Membrane Microdomains - ultrastructure, membrane proteins, Membrane Proteins - genetics, Membranes, Microscopy, Microscopy, Fluorescence - methods, Microscopy, Phase-Contrast - methods, Perceptual localization, polymers, Proteins, Spheroplasts, Spheroplasts - chemistry, Spheroplasts - metabolism, Spheroplasts - ultrastructure, Studies, Transferases (Other Substituted Phosphate Groups) - genetics, yellow fluorescent protein