Restoration of electron transport without proton pumping in mammalian mitochondria

被引：64

作者：

Perales-Clemente, Ester ^{[1
]}

Pilar Bayona-Bafaluy, Maria ^{[1
]}

Perez-Martos, Acisclo ^{[1
]}

Barrientos, Antoni ^{[2
,3
]}

Fernandez-Silva, Patricio ^{[1
]}

Antonio Enriquez, Jose ^{[1
]}

机构：

[1] Univ Zaragoza, Dept Bioquim, E-50013 Zaragoza, Spain

[2] Univ Miami, Miller Sch Med, Dr John T Macdonald Fdn Ctr Med Genet, Dept Neurol, Miami, FL 33136 USA

[3] Univ Miami, Miller Sch Med, Dr John T Macdonald Fdn Ctr Med Genet, Dept Biochem & Mol Biol, Miami, FL 33136 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2008年 / 105卷 / 48期

关键词：

AOX; mouse; oxidative phosphorylation; NDI1; CoQ;

D O I：

10.1073/pnas.0810518105

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We have restored the CoQ oxidative capacity of mouse mtDNA-less cells (p cells) by transforming them with the alternative oxidase Aox of Emericella nidulans. Cotransforming p cells with the NADH dehydrogenase of Saccharomyces cerevisiae, Ndi1 and Aox recovered the NADH DH/CoQ reductase and the CoQ oxidase activities. CoQ oxidation by AOX reduces the dependence of rho degrees cells on pyruvate and uridine. Coexpression of AOX and NDI1 further improves the recycling of NAD(+). Therefore, 2 single-protein enzymes restore the electron transport in mammalian mitochondria substituting >80 nuclear DNA-encoded and 11 mtDNA-encoded proteins. Because those enzymes do not pump protons, we were able to split electron transport and proton pumping (ATP synthesis) and inquire which of the metabolic deficiencies associated with the loss of oxidative phosphorylation should be attributed to each of the 2 processes.

引用

页码：18735 / 18739

页数：5

共 14 条

[1] Lack of complex I activity in human cells carrying a mutation in MtDNA-encoded ND4 subunit is corrected by the Saccharomyces cerevisiae NADH-quinone oxidoreductase (NDI1) gene [J].

Bai, YD ;

Hájek, P ;

Chomyn, A ;

Chan, E ;

Seo, BB ;

Matsuno-Yagi, A ;

Yagi, T ;

Attardi, G .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (42) :38808-38813

[2] Revisiting the mouse mitochondrial DNA sequence [J].

Bayona-Bafaluy, MP ;

Acín-Pérez, R ;

Mullikin, JC ;

Park, JS ;

Moreno-Loshuertos, R ;

Hu, PQ ;

Pérez-Martos, A ;

Fernández-Silva, P ;

Bai, YD ;

Enríquez, JA .

NUCLEIC ACIDS RESEARCH, 2003, 31 (18) :5349-5355

[3]

CHOMYN A, 1994, AM J HUM GENET, V54, P966

[4] Isolation of biogenetically competent mitochondria from mammalian tissues and cultured cells [J].

Fernández-Vizarra, E ;

López-Pérez, MJ ;

Enriquez, JA .

METHODS, 2002, 26 (04) :292-297

[5] Allotopic expression of a mitochondrial alternative oxidase confers cyanide resistance to human cell respiration [J].

Hakkaart, GAJ ;

Dassa, EP ;

Jacobs, HT ;

Rustin, P .

EMBO REPORTS, 2006, 7 (03) :341-345

[6]

Hofhaus G, 1996, Methods Enzymol, V264, P476, DOI 10.1016/S0076-6879(96)64043-9

[7] CYTOPLASMIC GENETICS OF MAMMALIAN-CELLS - CONDITIONAL SENSITIVITY TO MITOCHONDRIAL INHIBITORS AND ISOLATION OF NEW MUTANT PHENOTYPES [J].

HOWELL, N ;

SAGER, R .

SOMATIC CELL GENETICS, 1979, 5 (06) :833-845

[8] HUMAN-CELLS LACKING MTDNA - REPOPULATION WITH EXOGENOUS MITOCHONDRIA BY COMPLEMENTATION [J].

KING, MP ;

ATTARDI, G .

SCIENCE, 1989, 246 (4929) :500-503

[9] Kinetics of integrated electron transfer in the mitochondrial respiratory chain: random collisions vs. solid state electron channeling [J].

Lenaz, Giorgio ;

Genova, Maria Luisa .

AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2007, 292 (04) :C1221-C1239

[10] DEVELOPMENT AND CHARACTERIZATION OF CONTINUOUS AVIAN CELL-LINES DEPLETED OF MITOCHONDRIAL-DNA [J].

MORAIS, R ;

DESJARDINS, P ;

TURMEL, C ;

ZINKEWICHPEOTTI, K .

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY, 1988, 24 (07) :649-658

← 1 2 →