INFLUENCE OF RESPIRATION AND ATP HYDROLYSIS ON PROTON-ELECTROCHEMICAL GRADIENT ACROSS INNER MEMBRANE OF RAT-LIVER MITOCHONDRIA AS DETERMINED BY ION DISTRIBUTION
INFLUENCE OF RESPIRATION AND ATP HYDROLYSIS ON PROTON-ELECTROCHEMICAL GRADIENT ACROSS INNER MEMBRANE OF RAT-LIVER MITOCHONDRIA AS DETERMINED BY ION DISTRIBUTION
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DOI：
10.1111/j.1432-1033.1974.tb03899.x
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发表时间：
1974-01-01
期刊：
影响因子：
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通讯作者：
NICHOLLS, DG
中科院分区：
文献类型：
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作者：
NICHOLLS, DG
材料与方法用0.25M蔗糖差速离心法从大鼠肝脏中制备线粒体.在0.9ml容量的有机玻璃室中通过Clark型氧电极测定氧摄取。所有实验均在23 ℃下进行，孵育培养基的初始pH为7.2。双缩脲法测定蛋白质含量。寡霉素、鱼藤酮、琥珀酸钠、3-羟基丁酸钠、ATP和ADP购自Sigma Chemical Co.;来自Calbiochem的缬氨霉素，和来自放射化学中心（阿默舍姆，England）的所有同位素。FCCP是P.G.博士送的礼物。海特勒所有其他试剂均为市售最高纯度。
MATERIALS AND METHODSMitochondria were prepared from rat liver by differential centrifugation in 0.25 M sucrose. Oxygen uptake was determined by a Clark-type oxygen electrode in a perspex chamber of 0.9 ml capacity. All experiments were performed at 23" C, and the initial pH of the incubation media was 7.2. Protein was determined by the biuret method. Oligomycin, rotenone, sodium succinate, sodium~~-3-hydroxybutyrate, ATP and ADP were obtained from Sigma Chemical Co.; valinomycin from Calbiochem, and all isotopes from the Radiochemical Centre (Amersham, England). FCCP was a gift from Dr P. G. Heytler. All other reagents were of the highest purity commercially available.