Myasoedova K N, Tsuprun V L
N.N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.
FEBS Lett. 1993 Jul 5;325(3):251-4. doi: 10.1016/0014-5793(93)81083-c.
Purified cytochrome P-450LM4 was found to be monodisperse in 20% glycerol by analytical ultracentrifugation. Its S20,w value was quite similar to that of hexameric P-450LM2. At lower glycerol concentrations the P-450LM4 showed a tendency to aggregate. The P-450LM4 oligomers were immobilized on Ultrogel A4 under conditions allowing only one covalent link to the matrix per oligomer. In the presence of SDS, the oligomers dissociated leaving only 1/6th of the initial amount of bound protein on the matrix, suggesting that the purified P-450LM4 is a hexamer. This was confirmed by electron microscopy. The quaternary structure of the P-450LM4 was similar to that demonstrated earlier for P-450LM2.
通过分析超速离心法发现,纯化的细胞色素P-450LM4在20%甘油中呈单分散状态。其S20,w值与六聚体P-450LM2的S20,w值非常相似。在较低甘油浓度下,P-450LM4有聚集的倾向。P-450LM4寡聚体在仅允许每个寡聚体与基质形成一个共价连接的条件下固定在Ultrogel A4上。在十二烷基硫酸钠(SDS)存在的情况下,寡聚体解离,基质上仅留下初始结合蛋白量的1/6,这表明纯化的P-450LM4是一种六聚体。这一点通过电子显微镜得到了证实。P-450LM4的四级结构与先前证明的P-450LM2的四级结构相似。