Taubert Martin
Faculty of Biological Sciences, Institute of Biodiversity, Friedrich Schiller University Jena, Jena, Germany.
Methods Mol Biol. 2019;2046:57-69. doi: 10.1007/978-1-4939-9721-3_5.
Stable isotope probing combined with metaproteomics enables the detection and characterization of active key species in microbial populations under near-natural conditions, which greatly helps to understand the metabolic functions of complex microbial communities. This is achieved by providing growth substrates labeled with heavy isotopes such as C, which will be assimilated into microbial biomass. After subsequent extraction of proteins and proteolytic cleavage into peptides, the heavy isotope enrichment can be detected by high-resolution mass spectrometric analysis, and linked to the functional and taxonomic characterization of these biomarkers. Here we provide protocols for obtaining isotopically labeled proteins and for downstream SIP-metaproteomics analysis.
稳定同位素探测与元蛋白质组学相结合，能够在接近自然的条件下检测和表征微生物群体中的活性关键物种，这极大地有助于理解复杂微生物群落的代谢功能。这是通过提供用重同位素（如碳）标记的生长底物来实现的，这些底物会被同化到微生物生物量中。在随后提取蛋白质并将其酶解成肽段后，通过高分辨率质谱分析可以检测到重同位素富集，并将其与这些生物标志物的功能和分类特征联系起来。在此，我们提供了获取同位素标记蛋白质以及进行下游稳定同位素探测-元蛋白质组学分析的方案。
