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Yu, Qing; Liu, Xinyue; Keller, Mark P; Navarrete-Perea, Jose; Zhang, Tian; Fu, Sipei; Vaites, Laura P; Shuken, Steven R; Schmid, Ernst; Keele, Gregory R; Li, Jiaming; Huttlin, Edward L; Rashan, Edrees H; Simcox, Judith; Churchill, Gary A; Schweppe, Devin K; Attie, Alan D; Paulo, Joao A; Gygi, Steven P
Nature communications, 02/2023, Letnik: 14, Številka: 1Journal Article
Targeted proteomics enables hypothesis-driven research by measuring the cellular expression of protein cohorts related by function, disease, or class after perturbation. Here, we present a pathway-centric approach and an assay builder resource for targeting entire pathways of up to 200 proteins selected from >10,000 expressed proteins to directly measure their abundances, exploiting sample multiplexing to increase throughput by 16-fold. The strategy, termed GoDig, requires only a single-shot LC-MS analysis, ~1 µg combined peptide material, a list of up to 200 proteins, and real-time analytics to trigger simultaneous quantification of up to 16 samples for hundreds of analytes. We apply GoDig to quantify the impact of genetic variation on protein expression in mice fed a high-fat diet. We create several GoDig assays to quantify the expression of multiple protein families (kinases, lipid metabolism- and lipid droplet-associated proteins) across 480 fully-genotyped Diversity Outbred mice, revealing protein quantitative trait loci and establishing potential linkages between specific proteins and lipid homeostasis.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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in: SICRIS
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