Characterization of monoacylglycerol acyltransferase 2 inhibitors by a novel probe in binding assays

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• 作者：Zhengping Ma^a ; Hannguang J. Chao^b ; Huji Turdi^b ; Jon J. Hangeland^b ; Todd Friends^b ; Lisa M. Kopcho^c ; R. Michael Lawrence^b ; Dong Cheng^a ; ^{dong.cheng@bms.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：MGAT2 ; Enzyme inhibitor binding assay ; Enzyme inhibitor kinetics ; Acyltransferase ; Triglyceride synthesis
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 May 2016
• 年：2016
• 卷：501
• 期：Complete
• 页码：48-55
• 全文大小：1095 K

文摘

Monoacylglycerol acyltransferase 2 (MGAT2) is a membrane-bound lipid acyltransferase that catalyzes the formation of diacylglycerol using monoacylglycerol and fatty acyl CoA as substrates. MGAT2 is important for intestinal lipid absorption and is an emerging target for the treatment of metabolic diseases. In the current study, we identified and characterized four classes of novel MGAT2 inhibitors. We established both steady state and kinetic binding assay protocols using a novel radioligand, [³H]compound A. Diverse chemotypes of MGAT2 inhibitors were found to compete binding of [³H]compound A to MGAT2, indicating the broad utility of [³H]compound A for testing various classes of MGAT2 inhibitors. In the dynamic binding assays, the kinetic values of MGAT2 inhibitors such as K_on, K_off, and T_1/2 were systematically defined. Of particular value, the residence times of inhibitors on MGAT2 enzyme were derived. We believe that the identification of novel classes of MGAT2 inhibitors and the detailed kinetic characterization provide valuable information for the identification of superior candidates for in vivo animal and clinical studies. The current work using a chemical probe to define inhibitory kinetics can be broadly applied to other membrane-bound acyltransferases.