Traceless semisynthesis of a set of histone 3 species bearing specific lysine methylation marks.
Traceless semisynthesis of a set of histone 3 species bearing specific lysine methylation marks.
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DOI：
10.1002/cbic.201402313
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发表时间：
2014-09-22
期刊：
影响因子：
3.2
通讯作者：
Ruthenburg, Alexander J.
中科院分区：
文献类型：
--
作者：
Chen, Zhonglei;Grzybowski, Adrian T.;Ruthenburg, Alexander J.
关键词：
对组蛋白翻译后修饰和安装和去除它们的酶的功能的相当大的机制见解来自于修饰组蛋白的体外实验，通常嵌入在核小体中。我们报道了原生样组蛋白3 （H3）的第一次半合成，在79位含有三甲基赖氨酸和二甲基赖氨酸，在36位含有三甲基氨酸，以及K9和K27三甲基化物种更容易和无迹可寻的半合成。这些半合成在几毫克的尺度上是实用的，也可以通过标记的组合产生H3。尽管H3K36me3和H3K79me2/3的作用途径尚未完全确定，但每种修饰都具有不同的功能后果。为此，我们证明了我们的半合成组蛋白，当重组成核小体时，是无偏结合伴侣发现的有价值的亲和力试剂，并在核小体水平上将它们与甲基赖氨酸类似物（MLA）进行了比较。
Considerable mechanistic insight into the function of histone posttranslational modifications and the enzymes that install and remove them derives from in vitro experiments with modified histones, often embedded in nucleosomes. We report the first semisyntheses of native-like histone 3 (H3) bearing tri- and di- methyllysines at position 79 and trimethylsine at 36, as well as more facile and traceless semisyntheses of K9 and K27 trimethylated species. These semisyntheses are practical in multi-milligram scale and can also generate H3 with combinations of marks. Each of these modifications has distinct functional consequences, although the pathways by which H3K36me3 and H3K79me2/3 act have not been entirely mapped. To this end, we demonstrate that our semisynthetic histones, when reconstituted into nucleosomes, are valuable affinity reagents for unbiased binding-partner discovery, and compare them to their methyllysine analog (MLA) counterparts at the nucleosome level.
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