Blanck J, Rohde K, Behlke J, Jänig G R, Pfeil D, Ruckpaul K
Acta Biol Med Ger. 1979;38(2-3):399-408.
The complex monooxygenatic enzyme exhibits different functional behaviour at different integrational levels, thus indicating distinct organizational states. The aerobic NADPH reduction of microsomes, solubilized and reconstituted systems follows a biphasic kinetics, the two phases are attributed to associated state (cluster) and random cytochrome P-450 reduction. States of different cytochrome P-450/reductase ratio (associates) could not be differentiated in rate. Detergents (Triton N-101, cholate) are capable of disintegrating the system, at last only monophasic slow reduction is observed. The hydroxylation activity follows the respective reduction behaviour. Sedimentation analysis proves the distinct structural states. Reconstitution of the system can be achieved by means of detergent dilution as well as by combining the constituents. The activity of the reconstituted system depends on the composition of the phospholipids as well as on its organizational state. The reassociation of the solubilized enzyme system at nearly microsomal components stoichiometry (Triton N-101 dilution) proves to be thermodynamically governed leading to self-organization of the system without matrix prerequisite. Individual step rate constants of the reduction reaction and other system parameters are accessible by means of a model treatment of the disintegrated system. Further application to mixed kinetics systems is in progress.
这种复杂的单加氧酶在不同的整合水平上表现出不同的功能行为，从而表明存在不同的组织状态。微粒体、溶解和重组系统的需氧NADPH还原遵循双相动力学，这两个阶段归因于相关状态（簇）和随机细胞色素P-450还原。不同细胞色素P-450/还原酶比率（缔合体）的状态在速率上无法区分。去污剂（Triton N-101、胆酸盐）能够分解该系统，最终仅观察到单相缓慢还原。羟基化活性遵循各自的还原行为。沉降分析证明了不同的结构状态。该系统的重组可以通过去污剂稀释以及通过组合各成分来实现。重组系统的活性取决于磷脂的组成及其组织状态。已溶解的酶系统以接近微粒体成分化学计量比（Triton N-101稀释）进行的重新缔合被证明是由热力学控制的，导致系统在没有基质前提条件下的自组织。通过对分解系统的模型处理可以获得还原反应的各个步骤速率常数和其他系统参数。对混合动力学系统的进一步应用正在进行中。
