Academic Journal
Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis
| العنوان: | Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis |
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| المؤلفون: | Higgins, Anna J., Flynn, Alex J., Marconnet, Anaïs, Musgrove, Laura J., Postis, Vincent L. G., Lippiat, Jonathan D., Chung, Chun-wa, Ceska, Tom, Zoonens, Manuela, Sobott, Frank, Muench, Stephen P. |
| المصدر: | Higgins , A J , Flynn , A J , Marconnet , A , Musgrove , L J , Postis , V L G , Lippiat , J D , Chung , C , Ceska , T , Zoonens , M , Sobott , F & Muench , S P 2021 , ' Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis ' , Communications Biology , vol. 4 , 1337 . https://doi.org/10.1038/s42003-021-02834-3 |
| سنة النشر: | 2021 |
| المجموعة: | Discovery - University of Dundee Online Publications |
| مصطلحات موضوعية: | Cryoelectron microscopy, Membranes, /dk/atira/pure/subjectarea/asjc/2700/2701, name=Medicine (miscellaneous), /dk/atira/pure/subjectarea/asjc/1300/1300, name=General Biochemistry,Genetics and Molecular Biology, /dk/atira/pure/subjectarea/asjc/1100/1100, name=General Agricultural and Biological Sciences |
| الوصف: | Membrane proteins are essential for cellular growth, signalling and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them from the membrane creates challenges in their structural and functional study. Although amphipols have been very effective for single-particle electron cryo-microscopy (cryoEM) and mass spectrometry, they rely on initial detergent extraction before exchange into the amphipol environment. Therefore, circumventing this pre-requirement would be a big advantage. Here we use an alternative type of amphipol: a cycloalkane-modified amphiphile polymer (CyclAPol) to extract Escherichia coli AcrB directly from the membrane and demonstrate that the protein can be isolated in a one-step purification with the resultant cryoEM structure achieving 3.2 Å resolution. Together this work shows that cycloalkane amphipols provide a powerful approach for the study of membrane proteins, allowing native extraction and high-resolution structure determination by cryoEM. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | https://discovery.dundee.ac.uk/en/publications/2d860d60-688f-441b-ae11-c20ec297c680 |
| DOI: | 10.1038/s42003-021-02834-3 |
| الاتاحة: | https://discovery.dundee.ac.uk/en/publications/2d860d60-688f-441b-ae11-c20ec297c680 https://doi.org/10.1038/s42003-021-02834-3 https://discovery.dundee.ac.uk/ws/files/71419362/s42003_021_02834_3.pdf http://www.scopus.com/inward/record.url?scp=85119852881&partnerID=8YFLogxK |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.65770630 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1038/s42003-021-02834-3 |
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