Rosen P, Segal M, Pecht I
Eur J Biochem. 1981 Nov;120(2):339-44. doi: 10.1111/j.1432-1033.1981.tb05709.x.
The electron transfer equilibrium and kinetics between azurin from Alcaligenes faecalis and cytochrome c551 from Pseudomonas aeruginosa have been studied. The equilibrium constant K = ([Cyt(III)] . [Az(I)])/([Cyt(II)] . [Az(II))]) = 0.5 at 25 degrees C is about seven times smaller than that observed between the cytochrome c551 and the titrations confirmed a 43-mV difference between the mid-point potentials of +266 mV and +309 mV for the Alcaligenes and Pseudomonas azurins respectively. The kinetics of the reaction between Alcaligenes azurin and Pseudomonas cytochrome c551 were investigated by the temperature-jump chemical relaxation method. Only a single relaxation mode was observed throughout the range of concentrations and temperatures examined. Thus, the slow relaxation time observed in the reaction between P. aeruginosa azurin and cytochrome c551 is not observed with the Alcaligenes azurin. The simplest mechanism that can therefore be ascribed to the investigated system is: [formula: see text]. This scheme is similar to that proposed earlier for the reaction between P. aeruginosa azurin and cytochrome c551 but does not involve the conformational transition proposed for azurin. The specific rates for the electron transfer are still fast: 1.8 x 10(6) M-1 . s-1 and 3.0 x 10(6) M-1 . s-1 respectively at 25 degrees C.
已对粪产碱杆菌中的天青蛋白与铜绿假单胞菌中的细胞色素c551之间的电子转移平衡和动力学进行了研究。在25℃时,平衡常数K =([细胞色素(III)]·[天青蛋白(I)])/([细胞色素(II)]·[天青蛋白(II)])= 0.5,大约比在细胞色素c551之间观察到的平衡常数小7倍,并且滴定结果证实,粪产碱杆菌和铜绿假单胞菌天青蛋白的中点电位分别为+266 mV和+309 mV,两者相差43 mV。采用温度跃升化学弛豫法研究了粪产碱杆菌天青蛋白与铜绿假单胞菌细胞色素c551之间反应的动力学。在所研究的浓度和温度范围内,仅观察到单一的弛豫模式。因此,在铜绿假单胞菌天青蛋白与细胞色素c551反应中观察到的缓慢弛豫时间,在粪产碱杆菌天青蛋白反应中未观察到。因此,可归因于所研究体系的最简单机制是:[公式:见原文]。该方案与先前提出的铜绿假单胞菌天青蛋白与细胞色素c551之间反应的方案相似,但不涉及天青蛋白的构象转变。电子转移的具体速率仍然很快:在25℃时分别为1.8×10⁶ M⁻¹·s⁻¹和3.0×10⁶ M⁻¹·s⁻¹。