Efficient Protein Digestion at Elevated Temperature in the Presence of Sodium Dodecyl Sulfate and Calcium Ions for Membrane Proteomics.
Efficient Protein Digestion at Elevated Temperature in the Presence of Sodium Dodecyl Sulfate and Calcium Ions for Membrane Proteomics.
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在十二烷基硫酸钠和钙离子存在下，在高温下高效消化蛋白质，用于膜蛋白质组学。
DOI：
10.1021/acs.analchem.9b00484
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发表时间：
2019
影响因子：
7.4
通讯作者：
Loraine J
中科院分区：
文献类型：
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作者：
Loraine J
膜蛋白的重要性日益增加，激发了持续发展和改进的方法，强大的膜蛋白质组学。在这里，我们开发了一种非常简单和有效的方法膜蛋白消化使用离子洗涤剂，十二烷基硫酸钠（SDS），在高温下，胰蛋白酶通常是灭活的条件。我们的研究结果表明，胰蛋白酶可以通过钙离子和氯化钠的组合来稳定，这使得蛋白质在高温下在强离子洗涤剂如SDS的存在下消化。寻找胰蛋白酶的稳定化条件为洗涤剂在膜蛋白研究中的应用提供了新的机会。
The growing significance of membrane proteins inspires continuous development and improvement of methods for robust membrane proteomics. Here, we developed a very simple and efficient method for membrane protein digestion using an ionic detergent, sodium dodecyl sulfate (SDS), at high temperature, conditions where trypsin is normally inactivated. Our results suggest that trypsin can be stabilized by a combination of calcium ions and sodium chloride, which enables protein digestion at elevated temperature in the presence of strong ionic detergents such as SDS. Finding the conditions for stabilization of trypsin offers novel opportunities for the application of detergents for the investigation of membrane proteins.