Purification, characterization and crystallization of the F-ATPase from Paracoccus denitrificans
| العنوان: | Purification, characterization and crystallization of the F-ATPase from Paracoccus denitrificans |
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| المؤلفون: | Michael J.O. Wakelam, Ian M. Fearnley, John E. Walker, Martin G. Montgomery, Ian N. Watt, Qifeng Zhang, Edgar Morales-Ríos, Shujing Ding |
| المساهمون: | Montgomery, Martin [0000-0001-6142-9423], Wakelam, Michael [0000-0003-4059-9276], Walker, John [0000-0001-7929-2162], Apollo - University of Cambridge Repository |
| المصدر: | Open Biology, Vol 5, Iss 9 (2015) 'Open Biology ', vol: 5, pages: 150119-1-150119-9 (2015) Open Biology |
| بيانات النشر: | The Royal Society, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | crystallization, Protein subunit, Immunology, subunits, General Biochemistry, Genetics and Molecular Biology, Mass Spectrometry, chemistry.chemical_compound, Adenosine Triphosphate, Bacterial Proteins, F-ATPase, Cardiolipin, α-proteobacteria, lcsh:QH301-705.5, Research Articles, chemistry.chemical_classification, paracoccus denitrificans, biology, Research, General Neuroscience, Inhibitor protein, biology.organism_classification, Transmembrane protein, Protein Subunits, Proton-Translocating ATPases, Enzyme, chemistry, Biochemistry, lcsh:Biology (General), f-atpase, Electrophoresis, Polyacrylamide Gel, Paracoccus denitrificans, cardiolipin, Adenosine triphosphate, Protein Binding |
| الوصف: | The structures of F-ATPases have been determined predominantly with mitochondrial enzymes, but hitherto no F-ATPase has been crystallized intact. A high-resolution model of the bovine enzyme built up from separate sub-structures determined by X-ray crystallography contains about 85% of the entire complex, but it lacks a crucial region that provides a transmembrane proton pathway involved in the generation of the rotary mechanism that drives the synthesis of ATP. Here the isolation, characterization and crystallization of an integral F-ATPase complex from the α-proteobacterium Paracoccus denitrificans are described. Unlike many eubacterial F-ATPases, which can both synthesize and hydrolyse ATP, the P. denitrificans enzyme can only carry out the synthetic reaction. The mechanism of inhibition of its ATP hydrolytic activity involves a ζ inhibitor protein, which binds to the catalytic F₁-domain of the enzyme. The complex that has been crystallized, and the crystals themselves, contain the nine core proteins of the complete F-ATPase complex plus the ζ inhibitor protein. The formation of crystals depends upon the presence of bound bacterial cardiolipin and phospholipid molecules; when they were removed, the complex failed to crystallize. The experiments open the way to an atomic structure of an F-ATPase complex. his work was funded by the intramural programme of the Medical Research Council via MRC programme U105663150 to J.E.W., and by support from the Biotechnology and Biological Sciences Research Council to M.J.O.W. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 2046-2441 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fc8855ba6fc274759745418a28887d99 https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.150119 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....fc8855ba6fc274759745418a28887d99 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20462441 |
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