Horner L, Flemming H W
Biol Chem Hoppe Seyler. 1985 Mar;366(3):303-10.
The covalent fixation of the phosphinoyl residues in the active site of alpha-chymotrypsin is proved by the application of the fluorescent phosphinoyl fluorides 1 [( 5-(dimethylamino)-1-naphthyl]phenylphosphinoyl-fluoride) or 4 [(5-methoxy-1-naphthyl)phenyl-phosphinoylfluoride]. The differences in the rates of the phosphinoylation of alpha-chymotrypsin and "methyl-alpha-chymotrypsin" as compared to 1 agree with model reactions. In both enzymes the serine-OH in the active site is phosphinoylated. The non-fluorescent 4-nitrophenyl [5-(dimethylamino)-1-naphthyl]phenylphospinate (3) and the corresponding non-fluorescent 5-methoxynaphthyl derivative 5 inhibit alpha-chymotrypsin far more slowly than the corresponding fluorides 1 and 4. The phosphinoyl residues of the nitrophenyl esters 3 and 5 are covalently linked in a yield of 80% to the active site of the enzyme with evolution of fluorescence. 20% of the nitrophenyl ester inhibits the enzyme by adsorption.
通过应用荧光磷酰氟1[（5-（二甲基氨基）-1-萘基]苯基磷酰氟）或4[（5-甲氧基-1-萘基）苯基磷酰氟]，证实了α-胰凝乳蛋白酶活性位点中磷酰基残基的共价固定。与1相比，α-胰凝乳蛋白酶和“甲基-α-胰凝乳蛋白酶”的磷酰化速率差异与模型反应一致。在这两种酶中，活性位点的丝氨酸-OH都被磷酰化。非荧光的4-硝基苯基[5-（二甲基氨基）-1-萘基]苯基膦酸酯（3）和相应的非荧光5-甲氧基萘基衍生物5对α-胰凝乳蛋白酶的抑制作用比相应的氟化物1和4慢得多。硝基苯基酯3和5的磷酰基残基以80%的产率共价连接到酶的活性位点，并伴随荧光产生。20%的硝基苯基酯通过吸附抑制酶。
