Differential steady-state tyrosine phosphorylation of two oligomeric forms of mitochondrial F0F1ATPsynthase: a structural proteomic analysis |
| |
| Authors: | Di Pancrazio Francesca Bisetto Elena Alverdi Vera Mavelli Irene Esposito Gennaro Lippe Giovanna |
| |
| Affiliation: | Department of Biomedical Sciences and Technologies, University of Udine, MATI Center of Excellence, Udine, Italy. |
| |
| Abstract: | We investigated tyrosine phosphorylation of F(0)F(1)ATPsynthase 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 F(1)gamma 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 F(1) central stalk. |
| |
| Keywords: | Blue native (BN)‐PAGE Dimerization Mitochondrial F0F1ATPsynthase LC‐ESI/MS Phosphotyrosines |
| 本文献已被 PubMed 等数据库收录！ |
