Chemical methods for the simultaneous quantitation of metabolites and proteins from single cells.
Chemical methods for the simultaneous quantitation of metabolites and proteins from single cells.
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DOI：
10.1021/jacs.5b00944
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发表时间：
2015-04-01
影响因子：
15
通讯作者：
Heath JR
中科院分区：
文献类型：
--
作者：
Xue M;Wei W;Su Y;Kim J;Shin YS;Mai WX;Nathanson DA;Heath JR
我们描述了单细胞综合代谢和蛋白质组测定的化学方法。细胞内代谢物（包括葡萄糖摄取和其他 3 种物质）的定量测定被设计为具有荧光读数的表面竞争性结合测定。这使得能够集成到具有功能性蛋白质免疫测定的微阵列格式中，所有这些都被整合到单细胞条形码芯片（SCBC）的微室中。使用 SCBC，我们询问人源性胶质母细胞瘤癌细胞对 EGFR 抑制的反应。我们首次报告了细胞间代谢异质性以及代谢物-磷蛋白相关网络的基线和药物诱导的变化。
We describe chemical approaches for integrated metabolic and proteomic assays from single cells. Quantitative assays for intracellular metabolites, including glucose uptake and 3 other species, are designed as surface-competitive binding assays with fluorescence readouts. This enables integration into a micro-array format with functional protein immunoassays, all of which are incorporated into the microchambers of a single-cell barcode chip (SCBC). Using the SCBC, we interrogate the response of human-derived glioblastoma cancer cells to EGFR inhibition. We report, for the first time, on both the intercellular metabolic heterogeneity as well as the baseline and drug-induced changes in the metabolite-phosphoprotein correlation network.