Laboratoire Innovations technologiques pour la Détection et le Diagnostic (Li2D), Service de Pharmacologie et Immunoanalyse (SPI), CEA, INRAE, F-30207 Bagnols-sur-Cèze, France.
Environ Microbiol. 2020 Aug;22(8):2996-3004. doi: 10.1111/1462-2920.14975. Epub 2020 Mar 11.
Differential shotgun proteomics identifies proteins that discriminate between sets of samples based on differences in abundance. This methodology can be easily applied to study (i) specific microorganisms subjected to a variety of growth or stress conditions or (ii) different microorganisms sampled in the same condition. In microbiology, this comparison is particularly successful because differing microorganism phenotypes are explained by clearly altered abundances of key protein players. The extensive description and quantification of proteins from any given microorganism can be routinely obtained for several conditions within a few days by tandem mass spectrometry. Such protein-centred microbial molecular phenotyping is rich in information. However, well-designed experimental strategies, carefully parameterized analytical pipelines, and sound statistical approaches must be applied if the shotgun proteomic data are to be correctly interpreted. This minireview describes these key items for a quick molecular phenotyping based on label-free quantification shotgun proteomics.
差异鸟枪法蛋白质组学可根据丰度差异来区分样本集,从而识别出不同的蛋白质。这种方法可轻松应用于以下研究:(i)特定微生物在多种生长或应激条件下的研究;(ii)同一条件下不同微生物的采样研究。在微生物学中,这种比较特别成功,因为不同微生物表型的出现是由关键蛋白丰度的明显改变所解释的。通过串联质谱法,在几天内就可以常规地从几种条件下获得给定微生物的大量蛋白质的详细描述和定量。这种以蛋白质为中心的微生物分子表型分析富含信息。然而,如果要正确解释鸟枪法蛋白质组学数据,则必须应用精心设计的实验策略、精心参数化的分析流程和合理的统计方法。本综述描述了基于无标记定量鸟枪法蛋白质组学的快速分子表型分析的这些关键要素。