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Fullerene–Interfaced Porphyrin Ligand in Affinity Chromatography of Membrane Proteins

机译:富勒烯-交联的卟啉配体在膜蛋白的亲和色谱中

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摘要

To purify the mammalian cell membrane porphyrin binding proteins (PBP), an original affinity chromatographic technique is proposed. The method is based on the application of an agarose attached fullerene-porphyrin ligand connected to a polysaccharide gel matrix through the epoxy-cyclohexyl residue interface. A selective PBP-stationary phase coupling has been managed in a single column chromatographic run leading to a complete purification of the 17.6 kDa monomer protein from the rat myocardium mitochondria membranes. A synchronous pH and ionic strength linear gradients were used to separate this protein with a high specific affinity to the porphyrin K related structures from all non-specific sorption retained membrane proteins.
机译:为了纯化哺乳动物细胞膜卟啉结合蛋白(PBP),提出了一种原始的亲和层析技术。该方法基于施加通过琼脂糖连接的富勒烯-卟啉配体,该配体通过环氧-环己基残基界面连接至多糖凝胶基质。选择性PBP固定相耦合已在单柱色谱运行中进行了处理,从而从大鼠心肌线粒体膜中完全纯化了17.6 kDa的单体蛋白。使用同步的pH和离子强度线性梯度将该蛋白与卟啉K相关的结构与所有非特异性吸附保留的膜蛋白分离,并具有对卟啉K相关结构的高特异性亲和力。

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