Chourey Karuna, Hettich Robert L
Chemical Sciences Division, Oak Ridge National Lab, Oak Ridge, TN, USA.
Methods Mol Biol. 2018;1841:293-302. doi: 10.1007/978-1-4939-8695-8_20.
Soil metaproteomics is a rapidly developing and rather complex field aimed at understanding the functionalities of soil microbial communities. One of the main challenges of such an approach is the availability of a robust and efficient protocol to extract proteins from soil microbes inhabiting this complex matrix. The wide range of soil types and the innumerable variations in soil properties confound this experimental goal. Here we present a detergent based, heat-assisted cellular lysis method coupled with trichloroacetic acid (TCA) precipitation of soil microbial proteins that has been developed in our lab and found to be reasonably robust and unbiased in extracting microbial proteins from a broad range of soils for downstream mass spectrometric characterizations of microbial metabolic activities in natural ecosystems.
土壤宏蛋白质组学是一个快速发展且相当复杂的领域，旨在了解土壤微生物群落的功能。这种方法的主要挑战之一是要有一个强大而有效的方案，从栖息于这种复杂基质中的土壤微生物中提取蛋白质。土壤类型的广泛以及土壤性质的无数变化使这一实验目标变得复杂。在此，我们展示了一种基于去污剂的热辅助细胞裂解方法，该方法结合了三氯乙酸（TCA）沉淀土壤微生物蛋白质，这是我们实验室开发的方法，已发现其在从多种土壤中提取微生物蛋白质方面相当稳健且无偏差，可用于下游对自然生态系统中微生物代谢活动进行质谱表征。
