Rein H, Maricic A, Jänig G R, Vuk-Pavlovic S, Benko B, Ristau O, Ruckpaul K
Biochim Biophys Acta. 1976 Sep 28;446(1):325-30. doi: 10.1016/0005-2795(76)90123-9.
Cytochrome P-450 was solubilized from phenobarbital induced rabbit liver and purified by affinity chromatography. The longitudinal proton magnetic relaxation rates of this ferric, low-spin sample (as confirmed by ESR) in 20% glycerol aqueous solution are very large compared with low-spin methaemoglobin and myoglobin derivatives. Similarly high rates were measured in a deuterated solution using the aliphatic protons of glycerol as stereochemical markers, which strongly suggests that the haem iron in cytochrome P-450 is much more accessible to the solvent than in harmoglobin or myoglobin. Type I substate (Spasman) produced small but significant increases in NMR rates both in the H2O and in the 2H2O solution, while binding of aniline (Type II substrate) doubled the rates.
细胞色素P - 450从苯巴比妥诱导的兔肝脏中溶解出来，并通过亲和色谱法进行纯化。该铁（III）低自旋样品（经电子顺磁共振证实）在20%甘油水溶液中的纵向质子磁弛豫率与低自旋高铁血红蛋白和肌红蛋白衍生物相比非常大。在使用甘油的脂肪族质子作为立体化学标记的氘代溶液中也测量到了类似的高弛豫率，这强烈表明细胞色素P - 450中的血红素铁比血红蛋白或肌红蛋白中的血红素铁更容易被溶剂接近。I型底物（司可巴比妥）在H₂O和²H₂O溶液中均使核磁共振弛豫率有小但显著的增加，而苯胺（II型底物）的结合使弛豫率加倍。
