Identification of Acyl Protein Thioesterases 1 and 2 as the Cellular Targets of the Ras-Signaling Modulators Palmostatin B and M
Identification of Acyl Protein Thioesterases 1 and 2 as the Cellular Targets of the Ras-Signaling Modulators Palmostatin B and M
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DOI：
10.1002/anie.201102967
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发表时间：
2011-01-01
影响因子：
16.6
通讯作者：
Waldmann, Herbert
中科院分区：
文献类型：
--
作者：
Rusch, Marion;Zimmermann, Tobias J.;Waldmann, Herbert
S-棕榈酰化和S-法尼基化的信号转导H-和N-Ras GTP酶在细胞生长、分裂和分化中起重要作用，并且在黑色素瘤、白血病和膀胱癌、肝癌和肾癌中经常突变。[1-3]通过棕榈酰转移酶和硫酯酶的H-和N-Ras的动态S-棕榈酰化和S-脱棕榈酰化决定细胞中适当的Ras定位和信号传导。[4]通过用α-内酯酰基蛋白硫酯酶1（APT 1）抑制剂Palmostatin B[5]或Palmostatin M [6]（图1）抑制去棕榈酰化来干扰Ras-去/再酰化循环，扰乱H-和N-Ras精确的稳态定位，并导致全局Ras信号转导的下调。Palmostatin B不抑制与Ras信号传导潜在相关的几种磷酸二酯酶。[5]然而，仍不清楚APT 1是否是细胞中唯一的Ras-去棕榈酰化酶，以及帕莫司他丁B和M是否靶向与Ras循环相关的其他蛋白质，特别是相关同工酶APT 2。[7]由于帕莫司他汀是酰化试剂，并且与APT 1一样，通过活性位点亲核试剂的可逆酰化抑制其靶蛋白，[5]它们的化学性质为通过慢反应蛋白质组学进行基于活性的蛋白质组分析（ABPP）进行靶点鉴定提供了机会。[8-9]在ABPP中，具有平衡反应性的化学探针被用于基于酶催化机制来标记、分离和鉴定酶靶标。通常，采用酶活性位点中的亲核试剂与嵌入选择性探针中的亲电试剂的反应。以前，基于β-内酯的探针已成功用于细菌[10-11]植物[12]以及哺乳动物蛋白质组的分析。[13]通过反应性蛋白质组学促进靶分离的常用策略是生物正交连接，
The S-palmitoylated and S-farnesylated signal transducing H-and N-Ras GTPases play important roles in cell growth, division and differentiation and are frequently mutated in melanoma, leukemia and cancers of the bladder, liver and kidney.[1-3] Dynamic S-palmitoylation and S-depalmitoylation of H-and N-Ras by palmitoyl transferases and thioesterases determines proper Ras localization and signalling in cells.[4] Interference with the Ras-de/reacylation cycle by inhibition of depalmitoylation with the ß-lactone acyl protein thioesterase 1 (APT1) inhibitors Palmostatin B[5] or Palmostatin M [6](Figure 1) disturbs H-and N-Ras precise steady-state localization, and results in down regulation of global Ras signalling. Palmostatin B does not inhibit several phosphodiesterases with potential relevance to Ras signalling.[5] However, it remains unclear whether APT1 is the only Ras-depalmitoylating enzyme in cells and whether Palmostatin B and M target additional proteins relevant to the Ras cycle, in particular the related isoenzyme APT2.[7]Since the Palmostatins are acylating reagents and, as in the case of APT1, inhibit their target proteins through reversible acylation of active-site nucleophiles,[5] their chemical nature offers an opportunity for target identification by means of activity-based proteome profiling (ABPP) through slow reactive proteomics.[8-9] In ABPP, chemical probes with balanced reactivity are employed to tag, isolate and identify enzyme targets based on their mechanism of enzymatic catalysis. Typically, the reaction of a nucleophile in the enzyme active site with an electrophile embedded in a selective probe is employed. Previously, ß-lactone-based probes have successfully been used for profiling of bacterial [10-11] plant,[12] as well as mammalian proteomes.[13] A frequently applied strategy to facilitate target isolation by means of reactive proteomics is a bioorthogonal ligation of