Faculty of Biology, Technion-Israel Institute of Technology, Technion City, Haifa, Israel.
Faculty of Biology, Technion-Israel Institute of Technology, Technion City, Haifa, Israel.
Methods Enzymol. 2023;686:1-28. doi: 10.1016/bs.mie.2023.04.003. Epub 2023 May 18.
The field of N-terminomics has been advancing with the development of novel methods that provide a comprehensive and unbiased view of the N-terminome. Negative selection N-terminomics enables the identification of free and naturally modified protein N-termini. Here, we present a streamlined protocol that combines two negative selection N-terminomics methods, LATE and HYTANE, to increase N-terminome coverage by 1.5-fold compared to using a single methodology. Our protocol includes sample preparation and data analysis of both methods and can be applied to studying the N-terminome of diverse samples. The suggested approach enables researchers to achieve a more detailed and accurate understanding of the N-terminome.
N-端组学领域随着新型方法的发展而不断推进，这些方法为 N-端组学提供了全面和无偏的视角。负选择 N-端组学可以鉴定游离的和天然修饰的蛋白质 N-末端。在这里，我们提出了一个简化的方案，将两种负选择 N-端组学方法 LATE 和 HYTANE 相结合，与单一方法相比，将 N-端组学的覆盖率提高了 1.5 倍。我们的方案包括两种方法的样品制备和数据分析，可以应用于研究不同样本的 N-端组学。所建议的方法使研究人员能够更详细和准确地了解 N-端组学。
