Isolation and characterization of mitochondria from human B lymphoblastoid cell lines.
Mitochondria were isolated from detergent-treated Epstein-Barr virus-transformed human lymphocytes to examine their potential use in the study of the functional expression of genetic disorders of the respiratory chain. The increase of cytochrome c oxidase activity in the mitochondrial fraction indicated a 6-fold purification of intact mitochondria. Polarographic and spectrophotometric studies revealed that the isolated mitochondria were functionally well preserved. Furthermore, the isolated mitochondria supported an active in organello protein synthesis, which was dependent on the presence of a respiratory substrate generating ATP and was essentially abolished by chloramphenicol or by a specific respiratory chain inhibitor, such as antimycin. Thus, B lymphoblastoid cell lines constitute a valuable source of mitochondria to investigate mitochondrial functions in patients affected by respiratory chain disorders.
Bourgeron
T
T
Unité de Recherches sur les Handicaps Génétiques de l'Enfant, INSERM U 12, Hôpital des Enfants-Malades, Paris.
Chretien
D
D
Rötig
A
A
Munnich
A
A
Rustin
P
P
D000968:Antimycin A / Q000031:analogs & derivatives / Q000494:pharmacology; D001402:B-Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D002374:Catalase / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods; D002461:Cell Line, Transformed; D002701:Chloramphenicol / Q000494:pharmacology; D003902:Detergents; D003576:Electron Transport Complex IV / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010085:Oxidative Phosphorylation / Q000187:drug effects; D010101:Oxygen Consumption / Q000187:drug effects; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification
D003902:Detergents; D011506:Proteins; C032456:antimycin; D000968:Antimycin A; D002701:Chloramphenicol; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D009251:NADPH-Ferrihemoprotein Reductase; D003576:Electron Transport Complex IV