Abstract
We investigated tyrosine phosphorylation of F0F 1ATPsynthase using 3-D blue native (BN)-SDS-PAGE, a refinement of the electrophoretic analysis of mitochondrial complexes. Bovine heart mitochondria were detergent-solubilized and subjected to BN-PAGE. Bands of ATPsynthase monomer (Vmon) and dimer (Vdim) were excised and submitted to SDS-PAGE and immunoblotting. One protein corresponding to F1γ subunit was detected by anti-phosphotyrosine antibody in monomer but not in dimer. This was confirmed by MS peptide mapping. LC-ESI/MS analysis after 3-D SDS-PAGE demonstrated phosphotyrosine in fragment 43-54. NetPhos scores predicted the phosphorylated residue to be Tyr52, in a solvent-accessible loop at the foot of the F1 central stalk.
Original language | English (US) |
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Pages (from-to) | 921-926 |
Number of pages | 6 |
Journal | Proteomics |
Volume | 6 |
Issue number | 3 |
DOIs | |
State | Published - Feb 2006 |
Keywords
- Blue native (BN)-PAGE
- Dimerization
- LC-ESI/MS
- Mitochondrial FF ATPsynthase
- Phosphotyrosines
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology