Determination of coenzyme Q biosynthesis in cultured cells without the necessity for lipid extraction

被引：4

作者：

Córdoba-Pedregosa, MD ^{[1
]}

Villalba, JM ^{[1
]}

Alcaín, FJ ^{[1
]}

机构：

[1] Univ Cordoba, Fac Ciencias, Dept Biol Celular Fisiol & Immunol, Cordoba 14014, Spain

来源：

ANALYTICAL BIOCHEMISTRY | 2005年 / 336卷 / 01期

关键词：

D O I：

10.1016/j.ab.2004.09.031

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Ubiquinone (coenzyme Q; CoQ) is the only lipophilic antioxidant that is endogenously synthesized by all organisms. CoQ biosynthesis is determined in vitro by supplying a racholabeled precursor and, after lipid extraction and CoQ separation by thin-layer chromatography or high-performance liquid chromatography, the radioactivity present in the sample is quantified. In the rapid and simple method described here, we avoid the use of organic solvents by supplying 4-hydroxy-[U-C-14]benzoate as radiolabeled precursor and precipitating CoQ with trichloroacetic acid (TCA). After TCA precipitation, all radioactivity was present in the precipitate and CoQ was the only radiolabeled molecule detected. The radioactive material was then solubilized with NaOH and quantified in a scintillation counter. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：60 / 63

页数：4

共 13 条

[1] Silencing of ubiquinone biosynthesis genes extends life span in Caenorhabditis elegans [J].

Asencio, C ;

Rodríguez-Aguilera, JC ;

Ruiz-Ferrer, M ;

Vela, J ;

Navas, P .

FASEB JOURNAL, 2003, 17 (06) :1135-+

[2] A case of mitochondrial encephalomyopathy associated with a muscle coenzyme Q10 deficiency [J].

Boitier, E ;

Degoul, F ;

Desguerre, I ;

Charpentier, C ;

François, D ;

Ponsot, G ;

Diry, M ;

Rustin, P ;

Marsac, C .

JOURNAL OF THE NEUROLOGICAL SCIENCES, 1998, 156 (01) :41-46

[3]

CLARKE CF, 1991, J BIOL CHEM, V266, P16636

[4] Biochemical functions of coenzyme Q10 [J].

Crane, FL .

JOURNAL OF THE AMERICAN COLLEGE OF NUTRITION, 2001, 20 (06) :591-598

[5]

FORSGREN M, 2004, UNPUB BIOCH J

[6] UBIQUINONE BIOSYNTHESIS BY THE MICROSOMAL FRACTION FROM RAT-LIVER [J].

KALEN, A ;

NORLING, B ;

APPELKVIST, EL ;

DALLNER, G .

BIOCHIMICA ET BIOPHYSICA ACTA, 1987, 926 (01) :70-78

[7] Cerebellar ataxia and coenzyme Q10 deficiency [J].

Lamperti, C ;

Naini, A ;

Hirano, M ;

De Vivo, DC ;

Bertini, E ;

Servidei, S ;

Valeriani, M ;

Lynch, D ;

Banwell, B ;

Berg, M ;

Dubrovsky, T ;

Chiriboga, C ;

Angelini, C ;

Pegoraro, E ;

DiMauro, S .

NEUROLOGY, 2003, 60 (07) :1206-1208

[8]

MALTESE WA, 1985, J BIOL CHEM, V260, P1524

[9] Coenzyme Q10 supplementation provides mild symptomatic benefit in patients with Parkinson's disease [J].

Müller, T ;

Büttner, T ;

Gholipour, AF ;

Kuhn, W .

NEUROSCIENCE LETTERS, 2003, 341 (03) :201-204

[10] Quinone-responsive multiple respiratory-chain dysfunction due to widespread coenzyme Q10 deficiency [J].

Rötig, A ;

Appelkvist, EL ;

Geromel, V ;

Chretien, D ;

Kadhom, N ;

Edery, P ;

Lebideau, M ;

Dallner, G ;

Munnich, A ;

Ernster, L ;

Rustin, P .

LANCET, 2000, 356 (9227) :391-395

← 1 2 →