Differential steady-state tyrosine phosphorylation of two oligomeric forms of mitochondrial F0F1ATPsynthase

A structural proteomic analysis

Francesca Di Pancrazio, Elena Bisetto, Vera Alverdi, Irene Mavelli, Gennaro Esposito, Giovanna Lippe

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)921-926
Number of pages6
JournalProteomics
Volume6
Issue number3
DOIs
StatePublished - Feb 1 2006

Fingerprint

Phosphotyrosine
Phosphorylation
Dimers
Proteomics
Tyrosine
Native Polyacrylamide Gel Electrophoresis
Polyacrylamide Gel Electrophoresis
Monomers
Mitochondria
Detergents
Heart Mitochondria
Peptide Mapping
Immunoblotting
Peptides
Antibodies
Foot
Anti-Idiotypic Antibodies
Proteins

Keywords

  • Blue native (BN)-PAGE
  • Dimerization
  • LC-ESI/MS
  • Mitochondrial FF ATPsynthase
  • Phosphotyrosines

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Differential steady-state tyrosine phosphorylation of two oligomeric forms of mitochondrial F0F1ATPsynthase : A structural proteomic analysis. / Di Pancrazio, Francesca; Bisetto, Elena; Alverdi, Vera; Mavelli, Irene; Esposito, Gennaro; Lippe, Giovanna.

In: Proteomics, Vol. 6, No. 3, 01.02.2006, p. 921-926.

Research output: Contribution to journalArticle

Di Pancrazio, Francesca ; Bisetto, Elena ; Alverdi, Vera ; Mavelli, Irene ; Esposito, Gennaro ; Lippe, Giovanna. / Differential steady-state tyrosine phosphorylation of two oligomeric forms of mitochondrial F0F1ATPsynthase : A structural proteomic analysis. In: Proteomics. 2006 ; Vol. 6, No. 3. pp. 921-926.
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