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Purification, characterization and crystallization of the F-ATPase from

DOI: 10.1098/rsob.150119 DOI Help

Authors: Edgar Morales-rios (MRC Laboratory of Molecular Biology) , Ian Watt (MRC Mitochondrial Biology Unit) , Qifeng Zhang (The Babraham Institute) , Shujing Ding (The Medical Research Council Mitochondrial Biology Unit) , Ian M. Fearnley (The Medical Research Council Mitochondrial Biology Unit) , Martin G. Montgomery (The Medical Research Council Mitochondrial Biology Unit) , Michael J. O. Wakelam (The Babraham Institute) , John E. Walker (The Medical Research Council Mitochondrial Biology Unit)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Open Biology , VOL 5 (9)

State: Published (Approved)
Published: September 2015

Open Access Open Access

Abstract: 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.

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

Other Facilities: ESRF

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