文章基本信息

标题：An optimized method using peptide arrays for the identification of in vitro substrates of lysine methyltransferase enzymes
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作者：Elyn M. Rowe ; Kyle K. Biggar
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2018
卷号：5
页码：118-124
DOI：10.1016/j.mex.2018.01.012
语种：English
出版社：Elsevier
摘要：Graphical abstractDisplay OmittedAbstractWhile a number of post-translational modifications (PTM), such as phosphorylation and ubiquitination, have been extensively studied, lysine methylation is emerging as an important PTM with implications in a growing number of diverse cellular processes. To date, there are approximately 5000 identified methylation sites on non-histone proteins, and as the methyllysine proteome expands it becomes important to identify the lysine methyltransferase enzymes responsible for each methylation event. The use of peptide SPOT methylation assay has proven to be a useful in the identification and validation of novel substrates for lysine methyltransferase enzymes as it uses a weak beta emitter coupled with fluorography to detect methylation events. The method described in this paper provides improvements to the typical protocol for this assay, as a highly sensitive tritium assay can be developed with less radioactivity than previously described. This protocol provides an inexpensive alternative to weak beta signal enhancer sprays and washes for use in lysine methylation peptide SPOT arrays, and a simple open-source method for array quantification.
关键词：proteomics;non-histone methylation;SMYD3;MAP3K2;VEGFR1