Resonance Raman studies on protocatechuate 3,4-dioxygenase-inhibitor complexes.

Resonance Raman studies on protocatechuate 3,4-dioxygenase-inhibitor complexes.
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原儿茶酸 3,4-双加氧酶抑制剂复合物的共振拉曼研究。

DOI:
10.1021/bi00512a028
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发表时间:
1981
期刊:
影响因子:
2.9
通讯作者:
Epstein,RM
Epstein,RM
中科院分区:
生物学3区
文献类型:
--
作者:
QueJr,L;Epstein,RM

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材料和方法所有化学品均是商业获得的并且无需进一步纯化即可使用。 3HB和4HB中羟基的邻位和对位的氘化是通过在密封NMR管中在N 2 下在D 2Q 中进行碱催化交换在140℃下加热10小时来实现的。所得溶液的NMR谱显示>95%氘化。根据公开的程序(Fujisawa 等人,1972a;May 等人,1978)从铜绿假单胞菌(ATCC 23975)中制备 PCD。结晶酶的比活性为65或更好。稳态抑制动力学实验在 Gilson K-1C 测氧仪上于 25°C 下进行。
Materials and Methods All chemicals were obtained commercially and used without further purification. Deuteration of the positions ortho and para to the hydroxyl group in 3HB and 4HB was effected by base-catalyzed exchange in D2Q under N2 in a sealed NMR tube heated at 140 C for 10 h. The NMR spectra of the resulting solutions showed> 95% deuteration. PCD was prepared from Pseudomonas aeruginosa (ATCC 23975) according to published procedures (Fujisawa et al., 1972a; May et al., 1978). The crystalline enzyme had a specific activity of 65 or better. Steady-state inhibition kinetic experiments were performed on a Gilson K-1C oxygraph at 25 C.
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