The site-specific installation of methyl-lysine analogs into recombinant histones
The site-specific installation of methyl-lysine analogs into recombinant histones
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DOI：
10.1016/j.cell.2006.12.041
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发表时间：
2007-03-09
期刊：
影响因子：
64.5
通讯作者：
Shokat, Kevan M.
中科院分区：
文献类型：
--
作者：
Simon, Matthew D.;Chu, Feixia;Shokat, Kevan M.
组蛋白赖氨酸残基可以是单甲基化、二甲基化或三甲基化的。这些翻译后修饰调节效应蛋白的亲和力，也可能影响染色质结构，而不依赖于它们作为衔接子的作用。为了研究组蛋白赖氨酸甲基化，特别是在染色质的背景下，我们已经开发了一种化学方法来安装类似物的甲基赖氨酸到重组蛋白。这种方法允许快速生成大量的组蛋白，其中甲基化的位点和程度可以被指定。我们证明，这些甲基赖氨酸类似物（MLAs）是功能相似的天然同行。这些甲基化的组蛋白，被用来检查特定的赖氨酸甲基化效应蛋白的结合和核小体重塑率的影响。这种将位点特异性和程度特异性甲基化引入重组组蛋白的简单方法为研究赖氨酸甲基化影响染色质结构和功能的生化机制提供了有力的工具。
Histone lysine residues can be mono-, di-, or trimethylated. These posttranslational modifications regulate the affinity of effector proteins and may also impact chromatin structure independent of their role as adaptors. In order to study histone lysine methylation, particularly in the context of chromatin, we have developed a chemical approach to install analogs of methyl lysine into recombinant proteins. This approach allows for the rapid generation of large quantities of histones in which the site and degree of methylation can be specified. We demonstrate that these methyl-lysine analogs (MLAs) are functionally similar to their natural counterparts. These methylated histones, were used to examine the influence of specific lysine methylation on the binding of effecter proteins and the rates of nucleosme remodeling. This simple method of introducing site-specific and degree-specific methylation into recombinant histones provides a powerful tool to investigate the biochemical mechanisms by which lysine methylation influences chromatin structure and function.