F-0 MEMBRANE DOMAIN OF ATP SYNTHASE FROM BOVINE HEART-MITOCHONDRIA - PURIFICATION, SUBUNIT COMPOSITION, AND RECONSTITUTION WITH F1-ATPASE
UNSPECIFIED. (1994) F-0 MEMBRANE DOMAIN OF ATP SYNTHASE FROM BOVINE HEART-MITOCHONDRIA - PURIFICATION, SUBUNIT COMPOSITION, AND RECONSTITUTION WITH F1-ATPASE. BIOCHEMISTRY, 33 (25). pp. 7971-7978. ISSN 0006-2960Full text not available from this repository.
The F-o membrane domain of the F1Fo-ATP synthase complex has been purified from bovine heart mitochondria. The purification procedure involves the removal of peripheral membrane proteins, including F-1-ATPase, from submitochondrial particles with guanidine hydrochloride, followed by extraction of F-o and other membrane proteins from the stripped membranes in the presence of the detergent n-dodecyl beta-D-maltoside. F-o was then purified by ion-exchange and dye ligand chromatography in the presence of the same detergent. Approximately 15 mg of pure F-o was recovered from 1.8 g of mitochondrial membrane protein. The purified F-o is a complex of nine different polypeptides. They are subunits a, b, c, d, e, F-6, and A6L characterized before in F1Fo-ATPase preparations, and two new hitherto undetected subunits, named f and g. The sequences of subunits f and g have been determined. They are not related significantly to any known protein, but subunit f appears to contain a membrane-spanning alpha-helix. Proteins f and g are also present in approximately stoichiometric amounts in a highly purified preparation of intact F1Fo-ATPase, and so it is concluded that they are authentic subunits of the bovine enzyme with unknown functions. Dibutyltin 3-hydroxyflavone, an inhibitor of F1Fo-ATPase, also binds to the purified F-o in detergent and competes for binding with venturicidin. In the presence of F-1 and OSCP, the purified F-o was reassembled into the intact F1Fo-ATPase complex. Therefore, this procedure provides a relatively abundant source of pure and functional F-o that is suitable for structural analysis.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry|
|Journal or Publication Title:||BIOCHEMISTRY|
|Publisher:||AMER CHEMICAL SOC|
|Official Date:||28 June 1994|
|Number of Pages:||8|
|Page Range:||pp. 7971-7978|
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