Kim Ki-Bum, Lee Joong-Won, Lee Chang Seok, Kim Bong-Woo, Choo Hyo-Jung, Jung Soon-Young, Chi Sung-Gil, Yoon Young-Sil, Yoon Gyesoon, Ko Young-Gyu
School of Life Sciences and Biotechnology, Korea University, Anam-Dong, Sungbook-Ku, Seoul, Korea.
Proteomics. 2006 Apr;6(8):2444-53. doi: 10.1002/pmic.200500574.
In order to detect and identify ubiquitous lipid raft marker proteins, we isolated lipid rafts from different mouse organs, including the liver, lung, large brain, and kidney, and analyzed their proteins via 2-DE. Many protein spots were determined to be ubiquitous in all of the lipid rafts, and were annotated via LC and MS/MS. Twelve proteins were identified as ubiquitous raft proteins, and most of these were determined to be mitochondrial proteins, including mortalin, prohibitin, voltage-dependent anion channel, ATP synthase, NADH dehydrogenase, and ubiquinol-cytochrome c reductase. Via immunoblotting, these proteins were shown to exist in detergent-resistant lipid rafts prepared using different organ tissues. Since these oxidation-reduction respiratory chains and ATP synthase complex were detected in detergent-resistant lipid raft fractions which had been isolated from the plasma membrane but not from the mitochondria, and found in the cell surface when determined by immunofluoresence and immunohistochemistry, we conclude that plasma membrane lipid rafts might contain oxidation-reduction respiratory chains and ATP synthase complex.
为了检测和鉴定普遍存在的脂筏标记蛋白，我们从不同的小鼠器官（包括肝脏、肺、大脑和肾脏）中分离出脂筏，并通过二维电泳分析其蛋白质。许多蛋白点被确定在所有脂筏中普遍存在，并通过液相色谱和串联质谱进行注释。12种蛋白质被鉴定为普遍存在的脂筏蛋白，其中大多数被确定为线粒体蛋白，包括mortalin、抑制素、电压依赖性阴离子通道、ATP合酶、NADH脱氢酶和泛醇-细胞色素c还原酶。通过免疫印迹法，这些蛋白质显示存在于使用不同器官组织制备的抗去污剂脂筏中。由于这些氧化还原呼吸链和ATP合酶复合物在从质膜而非线粒体分离的抗去污剂脂筏组分中被检测到，并且通过免疫荧光和免疫组织化学测定发现它们存在于细胞表面，我们得出结论，质膜脂筏可能包含氧化还原呼吸链和ATP合酶复合物。
