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

被引：20

作者：

Bisetto, Elena ^{[1
,2
]}

Picotti, Paola ^{[1
,2
]}

Giorgio, Valentina ^{[1
,2
]}

Alverdi, Vera ^{[1
,2
]}

Mavelli, Irene ^{[1
,2
]}

Lippe, Giovanna ^{[1
,2
]}

机构：

[1] Univ Udine, Dept Biomed Sci & Technol, I-33100 Udine, Italy

[2] Univ Udine, MATI Ctr Excellence, I-33100 Udine, Italy

来源：

JOURNAL OF BIOENERGETICS AND BIOMEMBRANES | 2008年 / 40卷 / 04期

关键词：

Mammalian F(0)F(1)ATP synthase; Self-association; Subunit e stoichiometry; LC-MS; MS; AQUA peptides;

D O I：

10.1007/s10863-008-9183-5

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

The role of the integral inner membrane subunit e in self-association of F(0)F(1)ATP 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(0)F(1)ATP 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, gamma and e subunits, respectively. Accuracy of the method was demonstrated by confirming the 1:1 stoichiometry of subunits gamma and b. Altogether, the results indicate that the integrity of a unique copy of subunit e is essential for self-association of mammalian F(0)F(1)ATP synthase.

引用

页码：257 / 267

页数：11

共 45 条

[1] Coupling of rotation and catalysis in F1-ATPase revealed by single-molecule imaging and manipulation [J].

Adachi, Kengo ;

Oiwa, Kazuhiro ;

Nishizaka, Takayuki ;

Furuike, Shou ;

Noji, Hiroyuki ;

Itoh, Hiroyasu ;

Yoshida, Masasuke ;

Kinosita, Kazuhiko, Jr. .

CELL, 2007, 130 (02) :309-321

[2] Stoichiometry of subunit e in rat liver mitochondrial H+-ATP synthase and membrane topology of its putative Ca2+-dependent regulatory region [J].

Arakaki, N ;

Ueyama, Y ;

Hirose, M ;

Himeda, T ;

Shibata, H ;

Futaki, S ;

Kitagawa, K ;

Higuti, T .

BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 2001, 1504 (2-3) :220-228

[3] Yeast mitochondrial F1F0-ATP synthase exists as a dimer:: identification of three dimer-specific subunits [J].

Arnold, I ;

Pfeiffer, K ;

Neupert, W ;

Stuart, RA ;

Schägger, H .

EMBO JOURNAL, 1998, 17 (24) :7170-7178

[4] Yeast mitochondrial F1F0-ATPase: the novel subunit e is identical Tim11 [J].

Arnold, I ;

Bauer, MF ;

Brunner, M ;

Neupert, W ;

Stuart, RA .

FEBS LETTERS, 1997, 411 (2-3) :195-200

[5] The modulation in subunits e and g amounts of yeast ATP synthase modifies mitochondrial cristae morphology [J].

Arselin, G ;

Vaillier, J ;

Salin, B ;

Schaeffer, J ;

Giraud, MF ;

Dautant, A ;

Brèthes, D ;

Velours, J .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (39) :40392-40399

[6] The GxxxG motif of the transmembrane domain of subunit e is involved in the dimerization/oligomerization of the yeast ATP synthase complex in the mitochondrial membrane [J].

Arselin, G ;

Giraud, MF ;

Dautant, A ;

Vaillier, J ;

Brèthes, D ;

Coulary-Salin, B ;

Schaeffer, J ;

Velours, J .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2003, 270 (08) :1875-1884

[7] Absolute quantification of the G protein-coupled receptor rhodopsin by LC/MS/MS using proteolysis product peptides and synthetic peptide standards [J].

Barnidge, DR ;

Dratz, EA ;

Martin, T ;

Bonilla, LE ;

Moran, LB ;

Lindall, A .

ANALYTICAL CHEMISTRY, 2003, 75 (03) :445-451

[8] Membrane topography and near-neighbor relationships of the mitochondrial ATP synthase subunits e, f, and g [J].

Belogrudov, GI ;

Tomich, JM ;

Hatefi, Y .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (34) :20340-20345

[9] The proximal N-terminal amino acid residues are required for the coupling activity of the bovine heart mitochondrial factor B [J].

Belogrudov, Grigory I. .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2008, 473 (01) :76-87

[10] Mammalian ATPsynthase monomer versus dimer profiled by blue native PAGE and activity stain [J].

Bisetto, Elena ;

Di Pancrazio, Francesca ;

Simula, Maria Paola ;

Mavelli, Irene ;

Lippe, Giovanna .

ELECTROPHORESIS, 2007, 28 (18) :3178-3185

← 1 2 3 4 5 →