Evaluation of sulfatase-directed quinone methide traps for proteomics.
Evaluation of sulfatase-directed quinone methide traps for proteomics.
复制标题
用于蛋白质组学的硫酸酯酶导向的醌甲基化物陷阱的评估。
DOI：
10.1016/j.bmc.2011.04.044
复制
发表时间：
2012
影响因子：
3.5
通讯作者：
Sewald,Norbert
中科院分区：
文献类型：
--
作者：
Lenger,Janina;Schroder,Marius;Ennemann,EvaC;Muller,Benjamin;Wong,Chi-Huey;Noll,Thomas;Dierks,Thomas;Hanson,SarahR;Sewald,Norbert
硫酸盐酶水解裂解硫酸盐酯通过独特的催化醛，这是由翻译后氧化引入。为了分析健康和疾病中的活性硫酸酯酶，需要基于活性的蛋白质组学工具。本文采用基于活性的蛋白质组学探针（ABPPs）对针对磺化酶的醌（QM）诱捕器进行了评价。从对氟甲基苯基硫酸盐支架开始，酶促生成的qm陷阱可以灭活铜绿假单胞菌和肺炎克雷伯菌的细菌芳基硫酸酯酶，以及人类类固醇硫酸酯酶。然而，多种酶生成的QMs形成、扩散和非特异性标记纯化酶。在复杂的蛋白质组中，QM标记依赖于硫酸盐酶，但也是非特异性的。因此，氟甲基苯基硫酸盐是磺化酶的不良abpp。
Sulfatases hydrolytically cleave sulfate esters through a unique catalytic aldehyde, which is introduced by a posttranslational oxidation. To profile active sulfatases in health and disease, activity-based proteomic tools are needed. Herein, quinone methide (QM) traps directed against sulfatases are evaluated as activity-based proteomic probes (ABPPs). Starting from a p-fluoromethylphenyl sulfate scaffold, enzymatically generated QM-traps can inactivate bacterial aryl sulfatases from Pseudomonas aeruginosa and Klebsiella pneumoniae, and human steroid sulfatase. However, multiple enzyme-generated QMs form, diffuse, and non-specifically label purified enzyme. In complex proteomes, QM labeling is sulfatase-dependent but also non-specific. Thus, fluoromethylphenyl sulfates are poor ABPPs for sulfatases.