Activity-based probes that target functional subclasses of phospholipases in proteomes.

Activity-based probes that target functional subclasses of phospholipases in proteomes.
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基于活性的探针,针对蛋白质组中磷脂酶的功能亚类。

DOI:
10.1021/ja1000505
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发表时间:
2010-03-17
影响因子:
15
通讯作者:
Cravatt, Benjamin F.
Cravatt, Benjamin F.
中科院分区:
化学1区
文献类型:
--
作者:
Tully, Sarah E.;Cravatt, Benjamin F.

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磷脂酶是代谢细胞膜的磷脂组分并在关键脂质信号传导途径中起作用的一组大而多样的酶。新型磷脂酶的分子表征将受益于基于其独特的底物特异性和催化性质选择性靶向这些酶的化学探针。在这里,我们提出了一套基于活性的蛋白质分析(ABPP)探针,含有关键的识别和反应性元素,针对磷脂酶的丝氨酸水解酶超家族的合成和表征。我们表明,这些探针准确地报告的sn-1和sn-2底物特异性的磷脂酶在细胞和组织蛋白质组,包括sn-1选择性磷脂酶DDHD 1和钙依赖性转酰酶活性参与内源性大麻素的生物合成。我们预计,这些磷脂酶定向ABPP探针将促进新的脂质代谢酶的发现,并提供有价值的见解,他们的底物偏好。
Phospholipases are a large and diverse set of enzymes that metabolize the phospholipid components of cell membranes and function in key lipid signaling pathways. The molecular characterization of novel phospholipases would benefit from chemical probes that selectively target these enzymes based on their distinct substrate specificity and catalytic properties. Here, we present the synthesis and characterization of a set of activity-based protein profiling (ABPP) probes that contain key recognition and reactivity elements for targeting phospholipases of the serine hydrolase superfamily. We show that these probes accurately report on the sn-1 and sn-2 substrate specificities of phospholipases in cell and tissue proteomes, including the sn-1 selective phoshpolipase DDHD1 and a calcium-dependent transacylase activity implicated in endocannabinoid biosynthesis. We anticipate that these phospholipase-directed ABPP probes will facilitate the discovery of new lipid-metabolizing enzymes and provide valuable insights into their substrate preferences.
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