Discovery of Modulators of Adipocyte Physiology Using Fully Functionalized Fragments.
Discovery of Modulators of Adipocyte Physiology Using Fully Functionalized Fragments.
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使用全功能化片段发现脂肪细胞生理学调节剂。
DOI：
10.1007/978-1-4939-7847-2_9
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Saez,Enrique
中科院分区：
文献类型：
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作者：
Galmozzi,Andrea;Parker,ChristopherG;Kok,BernardP;Cravatt,BenjaminF;Saez,Enrique
与肥胖相关的脂肪细胞功能缺陷驱动全身性胰岛素抵抗和2型糖尿病的发展。纠正肥胖相关的脂肪细胞功能障碍的药物在人类中充当有用的胰岛素增敏剂，如噻唑烷二酮（TZD）所例示。我们开发了一个新的平台，将先进的化学蛋白质组学与表型筛选相结合，以识别促进脂肪细胞分化和脂质储存的小分子，以及它们的分子靶点。这些分子模拟TZDs在培养物中的活性，因此也可以在体内用作胰岛素增敏剂。该平台的核心是使用全功能化片段（FFF）探针，其由连接至炔-二氮杂环丙烯基团的可变片段样识别元件组成，所述识别元件能够直接在活细胞中光活化捕获探针结合的蛋白质，并随后将铜催化的叠氮化物-炔环加成至报告标签，以通过质谱法富集和鉴定这些探针结合的蛋白质。该平台可适用于脂肪细胞以外的多种筛选和细胞类型，有可能发现新的生物学途径，这些途径可用于影响人类疾病的药理学调节。
Defects in adipocyte function associated with obesity drive the development of systemic insulin resistance and type 2 diabetes. Agents that correct obesity-linked adipocyte dysfunction serve as useful insulin sensitizers in humans, as is exemplified by the thiazolidinediones (TZDs). We have developed a new platform that integrates advanced chemoproteomics with phenotypic screening to identify small molecules that promote differentiation and lipid storage in adipocytes, and, in tandem, their molecular target(s). These molecules mimic the activity of TZDs in culture and thus may also serve as insulin sensitizers in vivo. Central to this platform is the use of fully functionalized fragment (FFF) probes that consist of a variable, fragment-like recognition element linked to an alkyne-diazirine group that enables the photoactivated capture of probe-bound proteins directly in living cells and subsequent copper-catalyzed azide-alkyne cycloaddition to reporter tags for enrichment and identification of these probe-bound proteins by mass spectrometry. This platform, which can be adapted to diverse screens and cell types beyond adipocytes, has the potential to uncover new biological pathways amenable to pharmacological modulation that may impact human disease.