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

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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 F0F1ATP 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.

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References

  • Adachi K, Oiwa K, Nishizaka T, Furuike S, Noji H, Itoh H, Yoshida Y, Kinosita K (2007) Cell 130:309–321

    Article  CAS  Google Scholar 

  • Arakaki N, Ueyama Y, Hirose M, Himeda T, Shibata H, Futaki S, Kitagawa K, Higuti T (2001) Biochim. Biophys. Acta 1504:220–228

    Article  CAS  Google Scholar 

  • Arnold I, Bauer MF, Brunner M, Neupert W, Stuart RA (1997) FEBS Lett. 411:195–200

    Article  CAS  Google Scholar 

  • Arnold I, Pfeiffer K, Neupert W, Stuart RA, Schagger H (1998) EMBO J. 17:7170–7178

    Article  CAS  Google Scholar 

  • Arselin G, Giraud MF, Dautant A, Vaillier J, Brethes D, Coulary-Salin B, Schaeffer J, Velours J (2003) Eur. J. Biochem. 270:1875–1884

    Article  CAS  Google Scholar 

  • Arselin G, Vaillier J, Salin B, Schaeffer J, Giraud MF, Dautant A, Brèthes D, Velours J (2004) J Biol Chem. 279:40392–40399

    Article  CAS  Google Scholar 

  • Barnidge DR, Dratz EA, Martin T, Bonilla LE, Moran LB, Lindall A (2003) Anal. Chem. 75:445–451

    Article  Google Scholar 

  • Belogrudov GI (2008) Arch Biochem Biophys. 473:76–87

    CAS  Google Scholar 

  • Belogrudov GI, Tomich JM, Hatefi Y (1996) J. Biol. Chem. 271:20340–20345

    Article  CAS  Google Scholar 

  • Bisetto E, Di Pancrazio F, Simula MP, Mavelli I, Lippe G (2007) Electrophoresis 28:3178–3185

    Article  CAS  Google Scholar 

  • Bowler MW, Montgomery MG, Braig K, Leslie AGW, Walker JE (2007) J. Biol. 282:14238–14342

    CAS  Google Scholar 

  • Bustos D, Velours J (2005) J. Biol. Chem. 280:29004–29010

    Article  CAS  Google Scholar 

  • Campanella M, Casswell E, Chong S, Farah Z, Wieckowski MR, Abramov AY, Tinker A, Duchen MR (2008) Cell Metab. 8:13–25

    Article  CAS  Google Scholar 

  • Chen C, Safena AK, Simcoke WN, Garboczi DN, Pedersen PL, Ko YH (2006) J. Biol. Chem. 281:13777–13783

    Article  CAS  Google Scholar 

  • Cheng D, Hoogenraad CC, Rush J, Ramm E, Schlager MA, Duong DM, Xu P, Wijayawardana SR, Hanfelt J, Nakagawa T, Sheng M, Peng J (2006) Mol. Cell. Proteomics 5:1158–1170

    Article  CAS  Google Scholar 

  • Collinson IR, Runswick MJ, Buchanan SK, Fearnley IM, Skehel JM, van Raaij MJ, Griffiths DE, Walker EJ (1994) Biochemistry 33:7971–7978

    Article  CAS  Google Scholar 

  • de Souza GA, Godoy LM, Mann M (2006) Genome Biol. 7:R72

    Article  Google Scholar 

  • Di Pancrazio F, Mavelli I, Isola M, Losano G, Pagliaro P, Harris DA, Lippe G (2004) Biochim. Biophys. Acta 1659:52–62

    Article  CAS  Google Scholar 

  • Di Pancrazio F, Bisetto E, Alverdi V, Mavelli I, Esposito E, Lippe G (2006) Proteomics 6:921–926

    Article  CAS  Google Scholar 

  • Dickson VK, Silvester JA, Fearnley IM, Leslie AG, Walker JE (2006) EMBO J. 25:2911–2918

    Article  CAS  Google Scholar 

  • Dienhart M, Pfeiffer K, Schagger H, Stuart RA (2002) J. Biol. Chem. 277:39289–39295

    Article  CAS  Google Scholar 

  • Dudkina NV, Sunderhaus S, Braun HP, Boekema EJ (2006) FEBS Lett. 580:3427–3432

    Article  CAS  Google Scholar 

  • Elias JE, Gygi SP (2007) Nat Methods 4:207–214

    Article  CAS  Google Scholar 

  • Everard-Gigot V, Dunn CD, Dolan BM, Brunner S, Jensen RE, Stuart RA (2005) Eukaryot. Cell. 4:346–355

    Article  CAS  Google Scholar 

  • Ferguson SJ, Harris DA, Radda GK (1977) Biochem. J. 162:351–357

    CAS  Google Scholar 

  • Fronzes R, Weinmann T, Vaillier J, Velours J, Brethes D (2006) Biochemistry 45:6715–6723

    Article  CAS  Google Scholar 

  • Gallet PF, Zachowski A, Julien R, Fellmann P, Devaux PF, Maftah A (1999) Biochim. Biophys. Acta 1418:61–70

    Article  CAS  Google Scholar 

  • Garcìa JJ, Morales-Rìos E, Cortés-Hernandez P, Rodrìguez-Zavala JS (2006) Biochemistry 45:12695–12703

    Article  CAS  Google Scholar 

  • Gerber SA, Rush J, Stemman O, Kirschner MW, Gygi SP (2003) Proc. Natl. Acad. Sci. USA 100:6940–6945

    Article  CAS  Google Scholar 

  • Gerber SA, Kettenbach AN, Rush J, Gygi SP (2007) Methods Mol. Biol. 359:71–86

    Article  CAS  Google Scholar 

  • Hong S, Pedersen PL (2003) Proteins 51:155–161

    Article  CAS  Google Scholar 

  • Meyer B, Wittig I, Trifilieff E, Karas M, Schagger H (2007) Mol Cell Proteomics 6:1690–1699

    Article  CAS  Google Scholar 

  • Minauro-Sanmiguel F, Wilkens S, García JJ (2005) Proc Natl Acad Sci U S A. 102:12356–12358

    Article  CAS  Google Scholar 

  • Paumard P, Vaillier J, Coulary B, Schaeffer J, Soubannier V, Mueller DM, Brethes D, di Rago JP, Velours J (2002) EMBO J. 21:221–230

    Article  CAS  Google Scholar 

  • Rubinstein JL, Walker JE, Henderson R (2003) EMBO J. 22:6182–6192

    Article  CAS  Google Scholar 

  • Senior A (2007) Cell 130:220–221

    Article  CAS  Google Scholar 

  • Strauss M, Hofhaus G, Schroder R, Kuhlbrandt W (2008) EMBO J. 27:1154–1160

    Article  CAS  Google Scholar 

  • Thomas D, Bron P, Weimann T, Dautant A, Giraud MF, Paumard P, Salin B, Cavalier A, Velours J, Brethes D (2008) Biol Cell

  • Tomasetig L, Di Pancrazio F, Harris DA, Mavelli I, Lippe G (2002) Biochim. Biophys. Acta 1556:133–141

    Article  CAS  Google Scholar 

  • Walker EJ, Lutter R, Dupuis A, Runswick MJ (1991) Biochemistry 30:5369–5378

    Article  CAS  Google Scholar 

  • Wittig I, Schagger H (2005) Proteomics 5:4338–4346

    Article  CAS  Google Scholar 

  • Wittig I, Schägger H (2008) Biochim Biophys Acta. 1777:592–598

    Article  CAS  Google Scholar 

  • Wittig I, Braun HP, Schägger H (2006) Nature Protocols 1:418–428

    Article  CAS  Google Scholar 

  • Wittig I, Velours J, Stuart R, Schägger H (2008) Mol Cell Proteomics 5:995–1004

    Google Scholar 

  • Zerbetto E, Vergani L, Dabbeni-Sala F (1997) Electrophoresis 18:2059–2064

    Article  CAS  Google Scholar 

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Author notes

  1. Paola Picotti

    Present address: Institute of Molecular Systems Biology, ETH Zuerich, Wolfgang-Pauli Strasse 16, CH-8093, Zuerich, Switzerland

Authors and Affiliations

  1. Department of Biomedical Sciences and Technologies and M.A.T.I. Centre of Excellence, University of Udine, Piazzale Kolbe 4, I-33100, Udine, Italy

    Elena Bisetto, Paola Picotti, Valentina Giorgio, Vera Alverdi, Irene Mavelli & Giovanna Lippe

Authors
  1. Elena Bisetto
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  2. Paola Picotti
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  3. Valentina Giorgio
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  4. Vera Alverdi
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  5. Irene Mavelli
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  6. Giovanna Lippe
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Corresponding author

Correspondence to Giovanna Lippe.

Additional information

Elena Bisetto and Paola Picotti contributed equally to this work.

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Bisetto, E., Picotti, P., Giorgio, V. et al. Functional and stoichiometric analysis of subunit e in bovine heart mitochondrial F0F1ATP synthase. J Bioenerg Biomembr 40, 257–267 (2008). https://doi.org/10.1007/s10863-008-9183-5

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  • DOI: https://doi.org/10.1007/s10863-008-9183-5

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