Photoactive analogs of farnesyl pyrophosphate containing benzoylbenzoate esters: Synthesis and application to photoaffinity labeling of yeast protein farnesyltransferase
Photoactive analogs of farnesyl pyrophosphate containing benzoylbenzoate esters: Synthesis and application to photoaffinity labeling of yeast protein farnesyltransferase
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DOI：
10.1021/jo9602736
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发表时间：
1996-11-01
影响因子：
3.6
通讯作者：
Distefano, MD
中科院分区：
文献类型：
--
作者：
Gaon, I;Turek, TC;Distefano, MD
法尼基焦磷酸（FPP）参与众多细胞过程，包括与癌症相关的Ras蛋白转化突变体的异戊二烯化。光活性类似物可提供有关结合法尼基焦磷酸的酶活性位点的有用信息；然而，此类化合物的可用性极其有限。含有二苯甲酮部分的分子因其有用的光化学性质而成为有吸引力的光亲和标记试剂。在此，描述了两种化合物3a和3b的合成，它们含有对位和间位取代的苯甲酰苯甲酸酯。化合物3a和3b是酵母蛋白法尼基转移酶（PFTase）相对于FPP的竞争性抑制剂，其Ki值分别为910和380 nM。两种化合物在光解时都会使PFTase失活，导致酶活性多达44%的失活。在[P - 32]3a或[P - 32]3b存在下对PFTase进行光解会导致β亚基优先被标记，这表明该亚基参与异戊二烯基识别。这些化合物应该是研究以FPP为底物的酶的有价值的工具。
Farnesyl pyrophosphate (FPP) is involved in a large number of cellular processes including the prenylation of transforming mutants of Ras proteins implicated in cancer. Photoactive analogs could provide useful information about enzyme active sites that bind farnesyl pyrophosphate; however, the availability of such compounds is extremely limited. Molecules that incorporate benzophenone moieties are attractive photoaffinity labeling reagents because of their useful photochemical properties. Here, the syntheses of two compounds, 3a and 3b, containing para- and meta-substituted benzoylbenzoates are described. Compounds 3a and 3b are competitive inhibitors (with respect to FPP) of yeast protein farnesyltransferase (PFTase) with K-i values of 910 and 380 nM, respectively. Both compounds inactivate PFTase upon photolysis, resulting in as much as 44% inactivation of enzyme activity. Photolysis of PFTase in the presence of [P-32]3a or of [P-32]3b results in preferential labeling of the beta subunit, suggesting that this subunit is involved in prenyl group recognition. These compounds should be valuable tools for studying enzymes that utilize FPP as a substrate.