Noël H, Goswami T, Pande S V
Biochemistry. 1985 Aug 13;24(17):4504-9. doi: 10.1021/bi00338a003.
Carnitine acylcarnitine translocase has been solubilized from inverted inner membrane vesicles of rat liver mitochondria with octyl glucoside and reconstituted into asolectin liposomes. For both processes, optimization of the detergent to phospholipid ratio was found crucial for obtaining reconstitutively active liposomes. Reassembly of the solubilized carrier into asolectin liposomes was achieved either by the octyl glucoside dilution method or by Extracti-Gel D column chromatography. The reconstituted system catalyzed exchange diffusion of carnitine, exhibited the expected inhibitor and temperature sensitivity, and discriminated between stereoisomers of octanoylcarnitine. The activity of unidirectional import of carnitine was low compared to exchange diffusion. It showed high-temperature sensitivity and a loss of activity on prolonged sonication that was regained by an appropriate freeze-thaw step subsequently.
肉碱脂酰肉碱转位酶已用辛基葡糖苷从大鼠肝脏线粒体的内翻内膜囊泡中溶解出来，并重新组装到大豆卵磷脂脂质体中。对于这两个过程，发现优化去污剂与磷脂的比例对于获得具有重组活性的脂质体至关重要。通过辛基葡糖苷稀释法或通过Extracti-Gel D柱色谱法可将溶解的载体重新组装到大豆卵磷脂脂质体中。重组系统催化肉碱的交换扩散，表现出预期的抑制剂和温度敏感性，并能区分辛酰肉碱的立体异构体。与交换扩散相比，肉碱单向转运的活性较低。它表现出高温敏感性，长时间超声处理后活性丧失，随后通过适当的冻融步骤可恢复活性。
