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Insights from quantitative metaproteomics and protein-stable isotope probing into microbial ecology

机译:从定量元蛋白质组学和蛋白质稳定同位素探查微生物生态学的见解

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摘要

The recent development of metaproteomics has enabled the direct identification and quantification of expressed proteins from microbial communities in situ, without the need for microbial enrichment. This became possible by (1) significant increases in quality and quantity of metagenome data and by improvements of (2) accuracy and (3) sensitivity of modern mass spectrometers (MS). The identification of physiologically relevant enzymes can help to understand the role of specific species within a community or an ecological niche. Beside identification, relative and absolute quantitation is also crucial. We will review label-free and label-based methods of quantitation in MS-based proteome analysis and the contribution of quantitative proteome data to microbial ecology. Additionally, approaches of protein-based stable isotope probing (protein-SIP) for deciphering community structures are reviewed. Information on the species-specific metabolic activity can be obtained when substrates or nutrients are labeled with stable isotopes in a protein-SIP approach. The stable isotopes (13C, 15N, 36S) are incorporated into proteins and the rate of incorporation can be used for assessing the metabolic activity of the corresponding species. We will focus on the relevance of the metabolic and phylogenetic information retrieved with protein-SIP studies and for detecting and quantifying the carbon flux within microbial consortia. Furthermore, the combination of protein-SIP with established tools in microbial ecology such as other stable isotope probing techniques are discussed.
机译:元蛋白质组学的最新发展使得能够直接鉴定和定量来自微生物群落的表达蛋白,而无需进行微生物富集。 (1)大大提高了元基因组数据的质量和数量,以及(2)改进了现代质谱仪(MS)的准确性和(3)灵敏度,这成为可能。生理相关酶的鉴定可以帮助了解特定物种在社区或生态环境中的作用。除了鉴定,相对和绝对定量也很重要。我们将在基于MS的蛋白质组分析中回顾无标签和基于标签的定量方法,以及定量蛋白质组数据对微生物生态学的贡献。此外,审查了基于蛋白质的稳定同位素探测(protein-SIP)解密社区结构的方法。当以蛋白质-SIP方法用稳定的同位素标记底物或营养物质时,可以获得有关物种特异性代谢活性的信息。稳定同位素( 13 C, 15 N, 36 S)被掺入蛋白质中,掺入率可用于评估代谢相应物种的活性。我们将专注于通过蛋白质SIP研究获得的代谢和系统发育信息的相关性,以及检测和量化微生物群落中的碳通量。此外,还讨论了蛋白质SIP与微生物生态学中已建立的工具(例如其他稳定的同位素探测技术)的组合。

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