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

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