Protein and RNA profiles are highly informative when generating a comprehensive understanding of the dynamics of complex biological systems. Quantitative correlations between protein and RNA profiles are not always high, yet simultaneous acquisition of both profiles remains challenging, in part because of the limited availability of samples and the inconvenience of separately preparing protein and RNA fractions. In a previous study, protein, DNA, and RNA fractions were simultaneously prepared from the same sample using phenol-guanidinium isothiocyanate reagent (P/GTC), although the performance of P/GTC-extracts in proteogenomic analyses remains poorly understood. We therefore evaluated the performance of the P/GTC-extraction method in proteogenomic analyses using standard HEK293-F cells and human peripheral blood neutrophils. The latter cell type is renowned for its extreme vulnerability to protein/RNA degradation, reflecting high protease and RNase activities. Our data indicate that the P/GTC extraction method provides superior protein profiles from neutrophil and HEK293-F cell samples for simultaneous preparation of RNA and protein, as compared with those from conventional protein extraction methods. The P/GTC extraction method therefore provides a powerful and robust tool for a broad range of proteogenomic studies.

中文翻译：

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使用苯酚-胍基硫氰酸盐试剂对细胞裂解液进行蛋白质组学分析

当对复杂的生物系统的动力学有全面的了解时，蛋白质和RNA的概况是非常有用的。蛋白质和RNA谱图之间的定量相关性并不总是很高，但是同时获取这两个谱图仍然具有挑战性，部分原因是样品的供应有限以及单独制备蛋白质和RNA馏分的不便。在先前的研究中，虽然对蛋白质组学分析中P / GTC提取物的性能了解甚少，但使用异硫氰酸苯胍-胍盐（P / GTC）从同一样品中同时制备了蛋白质，DNA和RNA馏分。因此，我们使用标准的HEK293-F细胞和人类外周血中性粒细胞评估了蛋白质组学分析中P / GTC提取方法的性能。后一种细胞类型因其对蛋白质/ RNA降解的极端脆弱性而闻名，反映出高的蛋白酶和RNase活性。我们的数据表明，与传统的蛋白质提取方法相比，P / GTC提取方法可从嗜中性粒细胞和HEK293-F细胞样品中提供优异的蛋白质谱，用于同时制备RNA和蛋白质。因此，P / GTC提取方法为广泛的蛋白质组学研究提供了强大而强大的工具。与传统的蛋白质提取方法相比。因此，P / GTC提取方法为广泛的蛋白质组学研究提供了强大而强大的工具。与传统的蛋白质提取方法相比。因此，P / GTC提取方法为广泛的蛋白质组学研究提供了强大而强大的工具。