Tully Sarah E, Cravatt Benjamin F
Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
J Am Chem Soc. 2010 Mar 17;132(10):3264-5. doi: 10.1021/ja1000505.
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 on the basis of their distinct substrate specificities 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 phospholipase 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.
磷脂酶是一类庞大且多样的酶，它们代谢细胞膜的磷脂成分，并在关键的脂质信号通路中发挥作用。新型磷脂酶的分子特征鉴定将受益于基于其独特底物特异性和催化特性选择性靶向这些酶的化学探针。在此，我们展示了一组基于活性的蛋白质谱分析（ABPP）探针的合成与表征，这些探针包含用于靶向丝氨酸水解酶超家族磷脂酶的关键识别和反应性元件。我们表明，这些探针能够准确报告细胞和组织蛋白质组中磷脂酶的sn-1和sn-2底物特异性，包括sn-1选择性磷脂酶DDHD1以及与内源性大麻素生物合成相关的钙依赖性转酰基酶活性。我们预计，这些针对磷脂酶的ABPP探针将有助于发现新的脂质代谢酶，并为它们的底物偏好提供有价值的见解。
