Novel experimental strategy for high resolution AFM imaging of membrane-associated bacterial toxins
| العنوان: | Novel experimental strategy for high resolution AFM imaging of membrane-associated bacterial toxins |
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| المؤلفون: | Jielin Sun, Zhifeng Shao, Meng-lin Luo, Yi Shen, Daniel M. Czajkowsky |
| المصدر: | Journal of Shanghai Jiaotong University (Science). 19:569-573 |
| بيانات النشر: | Springer Science and Business Media LLC, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Experimental strategy, Multidisciplinary, Membrane, Materials science, Membrane protein, Atomic force microscopy, Bilayer, Resolution (electron density), High resolution, Nanotechnology, Lipid bilayer |
| الوصف: | Bacterial pore-forming toxins (PFTs) are essential virulence factors of many human pathogens. Knowledge of their structure within the membrane is critical for an understanding of their function in pathogenesis and for the development of useful therapy. Atomic force microscopy (AFM) has often been employed to structurally interrogate many membrane proteins, including PFTs, owing to its ability to produce sub-nanometer resolution images of samples under aqueous solution. However, an absolute prerequisite for AFM studies is that the samples are single-layered and closely-packed, which is frequently challenging with PFTs. Here, using the prototypical member of the cholesterol-dependent cytolysin family of PFTs, perfringolysin O (PFO), as a test sample, we have developed a simple, highly robust method that routinely produces clean, closely-packed samples across the entire specimen surface. In this approach, we first use a small Teflon well to prepare the supported lipid bilayer, remove the sample from the well, and then directly apply the proteins to the bilayer. For reasons that are not clear, bilayer preparation in the Teflon well is essential. We anticipate that this simple method will prove widely useful for the preparation of similar samples, and thereby enable AFM imaging of the greatest range of bacterial PFTs to the highest possible resolution. |
| تدمد: | 1995-8188 1007-1172 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::6958602f1da5eccfb762192d34d5359a https://doi.org/10.1007/s12204-014-1543-1 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi...........6958602f1da5eccfb762192d34d5359a |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 19958188 10071172 |
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