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The α‐subunit of the mitochondrial F 1 ATPase interacts directly with the assembly factor Atp12p
Author(s) -
Wang ZhenGuo,
Sheluho Dmitry,
Gatti Domenico L.,
Ackerman Sharon H.
Publication year - 2000
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/19.7.1486
Subject(s) - biology , protein subunit , g alpha subunit , alpha (finance) , atpase , microbiology and biotechnology , mitochondrion , genetics , biochemistry , enzyme , gene , medicine , construct validity , nursing , patient satisfaction
The Atp12p protein of Saccharomyces cerevisiae is required for the assembly of the F 1 component of the mitochondrial F 1 F 0 ATP synthase. In this report, we show that the F 1 α‐subunit co‐precipitates and co‐purifies with a tagged form of Atp12p adsorbed to affinity resins. Moreover, sedimentation analysis indicates that in the presence of the F 1 α‐subunit, Atp12p behaves as a particle of higher mass than is observed in the absence of the α‐subunit. Yeast two‐hybrid screens confirm the direct association of Atp12p with the α‐subunit and indicate that the binding site for the assembly factor lies in the nucleotide‐binding domain of the α‐subunit, between Asp133 and Leu322. These studies provide the basis for a model of F 1 assembly in which Atp12p is released from the α‐subunit in exchange for a β‐subunit to form the interface that contains the non‐catalytic adenine nucleotide‐binding site.