Inhibitors of the isoprenylated protein endoprotease.
Inhibitors of the isoprenylated protein endoprotease.
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异戊二烯化蛋白质内切蛋白酶的抑制剂。
DOI：
10.1021/bi00060a033
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Rando,RR
中科院分区：
文献类型：
--
作者：
Ma,YT;Gilbert,BA;Rando,RR
异戊二烯化途径需要内切蛋白酶在异戊二烯化的半胱氨酸残基处切割修饰的蛋白质。这种内切蛋白酶很容易用简单的7V-乙酰基-5 ′-法尼基-L-Cys-（AFC）-Val-Ile-Met型四肽底物进行测定，其中AFC和三肽是水解产物。内切蛋白酶被证明不受（1）丝氨酸蛋白酶抑制剂，包括（2）半胱氨酸蛋白酶抑制剂，包括E-64和亮抑酶肽[然而，该酶被p-（羟汞）苯甲酸盐]，（3）金属蛋白酶抑制剂，包括磷酰胺、EDTA和1，10-菲咯啉，或（4）乙酰基蛋白酶抑制剂胃蛋白酶抑制剂。从这些数据得出的结论是，该酶可能不是金属酶。含有他汀部分的A-Boc-Sa/Z-反式-法尼基-L-半胱氨酸（BFC）衍生物也不是抑制性的，强烈表明该酶不是乙酰基蛋白酶。然而，该酶被BFC的醛衍生物（= 1.9 µ）有效抑制，这与该酶是丝氨酸或半胱氨酸蛋白酶的想法一致。制备了有效的基于四肽的竞争性抑制剂。类似物与易裂键修改，使水解不能发生是优秀的抑制剂。
The isoprenylation pathway requires an endoprotease that cleaves the modified protein at the isoprenylated cysteine residue. This endoprotease was readily assayed with simple tetrapeptide substrates of the type 7V-acetyl-5'-farnesyl-L-Cys-(AFC)-Val-Ile-Met, where AFC and the tripeptide are the products of the hydrolysis. The endoprotease proved to be unaffected by (1) serine protease inhibitors, including (4-amidinophenyl) methanesulfonyl fluoride, aprotinin, and leupeptin, by (2) cysteineprotease inhibitors, including E-64 and leupeptin [the enzyme is, however, inhibited by p-(hydroxymercuri) benzoate], by (3) metalloprotease inhibitors, including phosphoramidon, EDTA, and 1, 10-phenanthroline, or by (4) the aspartyl protease inhibitor pepstatin. The conclusion from these data is that the enzyme is probably not a metalloenzyme. A-Boc-Sa/Z-irani-farnesyl-L-cysteine (BFC) derivatives containing a statine moiety are also not inhibitory, strongly suggesting that the enzyme is not an aspartyl protease. However, the enzyme is potently inhibited by the aldehyde derivative of BFC (= 1.9 µ), which is consistent with the idea that the enzyme is a serine or cysteine protease. Potent tetrapeptide-based competitiveinhibitors were prepared. Analogs with the scissile bond modified so that hydrolysis could not occur were excellent inhibitors.