The role of the integral inner membrane subunit e in self-association of F0F1ATP synthase from bovine heart mitochondria was analyzed by in situ limited proteolysis, blue native PAGE/iterative SDS-PAGE, and LC-MS/MS. Selective degradation of subunit e, without disrupting membrane integrity or ATPase capacity, altered the oligomeric distribution of F 0F1ATP synthase, by eliminating oligomers and reducing dimers in favor of monomers. The stoichiometry of subunit e was determined by a quantitative MS-based proteomics approach, using synthetic isotope-labelled reference peptides IAQL*EEVK, VYGVGSL*ALYEK, and ELAEAQEDTIL*K to quantify the b, γ and e subunits, respectively. Accuracy of the method was demonstrated by confirming the 1:1 stoichiometry of subunits γ and b. Altogether, the results indicate that the integrity of a unique copy of subunit e is essential for self-association of mammalian F0F1ATP synthase. © 2008 Springer Science+Business Media, LLC.

Functional and stoichiometric analysis of subunit e in bovine heart mitochondrial F0F1ATP synthase / Bisetto E.; Picotti P.; Giorgio V.; Alverdi V.; Mavelli I.; Lippe G.. - In: JOURNAL OF BIOENERGETICS AND BIOMEMBRANES. - ISSN 0145-479X. - ELETTRONICO. - 40:4(2008), pp. 257-267. [10.1007/s10863-008-9183-5]

Functional and stoichiometric analysis of subunit e in bovine heart mitochondrial F0F1ATP synthase

Giorgio V.;Mavelli I.;
2008

Abstract

The role of the integral inner membrane subunit e in self-association of F0F1ATP synthase from bovine heart mitochondria was analyzed by in situ limited proteolysis, blue native PAGE/iterative SDS-PAGE, and LC-MS/MS. Selective degradation of subunit e, without disrupting membrane integrity or ATPase capacity, altered the oligomeric distribution of F 0F1ATP synthase, by eliminating oligomers and reducing dimers in favor of monomers. The stoichiometry of subunit e was determined by a quantitative MS-based proteomics approach, using synthetic isotope-labelled reference peptides IAQL*EEVK, VYGVGSL*ALYEK, and ELAEAQEDTIL*K to quantify the b, γ and e subunits, respectively. Accuracy of the method was demonstrated by confirming the 1:1 stoichiometry of subunits γ and b. Altogether, the results indicate that the integrity of a unique copy of subunit e is essential for self-association of mammalian F0F1ATP synthase. © 2008 Springer Science+Business Media, LLC.
2008
Functional and stoichiometric analysis of subunit e in bovine heart mitochondrial F0F1ATP synthase / Bisetto E.; Picotti P.; Giorgio V.; Alverdi V.; Mavelli I.; Lippe G.. - In: JOURNAL OF BIOENERGETICS AND BIOMEMBRANES. - ISSN 0145-479X. - ELETTRONICO. - 40:4(2008), pp. 257-267. [10.1007/s10863-008-9183-5]
Bisetto E.; Picotti P.; Giorgio V.; Alverdi V.; Mavelli I.; Lippe G.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/794367
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