Galmozzi Andrea, Dominguez Eduardo, Cravatt Benjamin F, Saez Enrique
Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
Methods Enzymol. 2014;538:151-69. doi: 10.1016/B978-0-12-800280-3.00009-8.
Activity-based protein profiling (ABPP) is a chemical proteomics approach that utilizes small-molecule probes to determine the functional state of enzymes directly in native systems. ABPP probes selectively label active enzymes, but not their inactive forms, facilitating the characterization of changes in enzyme activity that occur without alterations in protein levels. ABPP can be a tool superior to conventional gene expression and proteomic profiling methods to discover new enzymes active in adipocytes and to detect differences in the activity of characterized enzymes that may be associated with disorders of adipose tissue function. ABPP probes have been developed that react selectively with most members of specific enzyme classes. Here, using as an example the serine hydrolase family that includes many enzymes with critical roles in adipocyte physiology, we describe methods to apply ABPP analysis to the study of adipocyte enzymatic pathways.
基于活性的蛋白质谱分析（ABPP）是一种化学蛋白质组学方法，它利用小分子探针直接在天然系统中确定酶的功能状态。ABPP探针选择性地标记活性酶，而不标记其无活性形式，有助于表征在蛋白质水平未发生改变的情况下酶活性的变化。ABPP可能是一种优于传统基因表达和蛋白质组学分析方法的工具，可用于发现脂肪细胞中具有活性的新酶，并检测可能与脂肪组织功能紊乱相关的已鉴定酶的活性差异。目前已开发出能与特定酶类的大多数成员选择性反应的ABPP探针。在此，我们以丝氨酸水解酶家族为例，该家族包含许多在脂肪细胞生理学中起关键作用的酶，描述将ABPP分析应用于脂肪细胞酶促途径研究的方法。
