Suspension Trapping (S-Trap) Is Compatible with Typical Protein Extraction Buffers and Detergents for Bottom-Up Proteomics
Suspension Trapping (S-Trap) Is Compatible with Typical Protein Extraction Buffers and Detergents for Bottom-Up Proteomics
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DOI：
10.1021/acs.jproteome.8b00891
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发表时间：
2019-03-01
影响因子：
4.4
通讯作者：
Levin, Yishai
中科院分区：
文献类型：
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作者：
Elinger, Dalia;Gabashvili, Alexandra;Levin, Yishai
自下而上的蛋白质组学对细胞和组织的分析从裂解开始，然后是溶液内消化。通常使用的裂解缓冲液包括洗涤剂和其他试剂，以实现有效的蛋白质溶解。然而，在大多数情况下，这些试剂与下游分析仪器不相容。最近介绍了一种溶解和清除方法，称为悬浮捕集（S-Trap）。我们评估了常用的裂解缓冲液与S-Trap的相容性：SDS、尿素、NP-40、RIPA和SDS与DTT （SDT）的相容性。我们发现S-Trap与所有被测试的缓冲兼容，SDS和SDT表现最好。在这些数据的基础上，我们预计该方法将改变基于质谱的蛋白质组学的实验计划，使其更加灵活和耐受各种裂解缓冲。质谱蛋白质组学数据已通过PRIDE合作伙伴存储库存入ProteomeXchange Consortium，数据集标识符为PXDO11665。
The analysis of cells and tissue by bottom-up proteomics starts with lysis, followed by in-solution digestion. Lysis buffers commonly used include detergents and other reagents for achieving efficient protein solubility. However, these reagents are, for the most part, incompatible with downstream analytical instrumentation. One method for in-solution digestion and cleanup, termed suspension trapping (S-Trap), has been recently introduced. We present an evaluation of the compatibility of commonly used lysis buffers with S-Trap: SDS, urea, NP-40, RIPA, and SDS with DTT (SDT). We show that S-Trap is compatible with all of the tested buffers, with SDS and SDT performing the best. On the basis of these data, we anticipate that the method will transform experimental planning for mass-spectrometry-based proteomics, making it far more flexible and tolerable of various lysis buffers. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXDO11665.