Sagami H, Korenaga T, Ogura K, Steiger A, Pyun H J, Coates R M
Institute for Chemical Reaction Science, Tohoku University, Sendai, Japan.
Arch Biochem Biophys. 1992 Sep;297(2):314-20. doi: 10.1016/0003-9861(92)90678-p.
Geranylgeranyl diphosphate synthase from rat liver was separated from farnesyl diphosphate synthase, the most abundant and widely occurring prenyltransferase, by DEAE-Toyopearl column chromatography. The enzyme catalyzed the formation of E,E,E-geranylgeranyl diphosphate (V) from isopentenyl diphosphate (II) and dimethylallyl diphosphate (I), geranyl diphosphate (III), or farnesyl diphosphate (IV) with relative velocities of 0.09:0.15:1. 3-Azageranylgeranyl diphosphate (VII), designed as a transition-state analog for the geranylgeranyl diphosphate synthase reaction, was synthesized and found to act as a specific inhibitor for this synthase, but not for farnesyl diphosphate synthase. Diphosphate V and its Z,E,E-isomer (VI) also inhibited geranylgeranyl diphosphate synthase, but the effect was not as striking as that of the aza analog VII. Specific inhibition of geranylgeranyl diphosphate synthase by VII was also observed in experiments with 100,000g supernatants of rat brain and liver homogenates which contained isopentenyl diphosphate isomerase and prenyltransferases including farnesyl diphosphate synthase as well as geranylgeranyl diphosphate synthase. For farnesyl:protein transferase from rat brain, however, the aza compound did not show a stronger inhibitory effect than E,E,E-geranylgeranyl diphosphate.
通过DEAE - Toyopearl柱色谱法，从大鼠肝脏中分离出香叶基香叶基二磷酸合酶，该酶与法尼基二磷酸合酶分离，法尼基二磷酸合酶是最丰富且广泛存在的异戊二烯基转移酶。该酶催化异戊烯基二磷酸（II）与二甲基烯丙基二磷酸（I）、香叶基二磷酸（III）或法尼基二磷酸（IV）形成E,E,E - 香叶基香叶基二磷酸（V），相对速度为0.09:0.15:1。设计了一种作为香叶基香叶基二磷酸合酶反应过渡态类似物的3 - 氮杂香叶基香叶基二磷酸（VII），并发现其可作为该合酶的特异性抑制剂，但对法尼基二磷酸合酶无抑制作用。二磷酸V及其Z,E,E - 异构体（VI）也能抑制香叶基香叶基二磷酸合酶，但其效果不如氮杂类似物VII显著。在含有异戊烯基二磷酸异构酶和包括法尼基二磷酸合酶以及香叶基香叶基二磷酸合酶在内的异戊二烯基转移酶的大鼠脑和肝匀浆经100,000g离心后的上清液实验中，也观察到VII对香叶基香叶基二磷酸合酶的特异性抑制作用。然而，对于大鼠脑来源的法尼基：蛋白质转移酶，该氮杂化合物并未表现出比E,E,E - 香叶基香叶基二磷酸更强的抑制作用。
