Rice Histone Propionylation and Generation of Chemically Derivatized Synthetic H3 and H4 Peptides for Identification of Acetylation Sites and Quantification.
Rice Histone Propionylation and Generation of Chemically Derivatized Synthetic H3 and H4 Peptides for Identification of Acetylation Sites and Quantification.
复制标题
水稻组蛋白丙酰化和化学衍生化合成 H3 和 H4 肽的生成，用于乙酰化位点的鉴定和定量。
DOI：
10.1007/978-1-0716-0179-2_6
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Saze H.
中科院分区：
文献类型：
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作者：
Espinas NA;Villar-Briones A;Roy MC;Saze H.
组蛋白在染色质动力学的研究中起着至关重要的作用，因为它们在基因表达调控中具有广泛的意义。组蛋白的修饰与相关蛋白质如转录因子和辅激活因子一起参与这些调控过程。可用于增强组蛋白分析的生化技术之一是使用丙酸酐的化学衍生化。在这个协议中，我们描述了使用丙酰化有效地衍生化酸提取组蛋白从大米。我们还合成了H3和H4胰蛋白酶肽，从而模拟衍生提取肽的性质，以帮助使用靶向质谱法进行鉴定和定量。在这里，我们提供了前体离子的质量和每个合成肽的保留时间（RT）。这些提供了有用的信息，以促进组蛋白数据分析。最后，我们注意到，我们将以纳摩尔（nM）浓度将这些合成肽分发给那些希望利用它们进行测定和进一步实验研究的人。
Histone proteins are crucial in the study of chromatin dynamics owing to their wide-ranging implications in the regulation of gene expression. Modifications of histones are integral to these regulatory processes in concert with associated proteins, such as transcription factors and coactivators. One of the biochemical techniques available to enhance analysis of histone proteins is chemical derivatization using propionic anhydride. In this protocol, we describe the use of propionylation to efficiently derivatize acid-extracted histones from rice. We also synthesize H3 and H4 tryptic peptides, thus mimicking the nature of derivatized extracted peptides to aid in identification and quantification using targeted-mass spectrometry. Here we make available the masses of the precursor ions and the retention times (RT) of each synthesized peptide. These provide useful information to facilitate histone data analysis. Lastly, we note that we will distribute these synthetic peptides in nanomolar (nM) concentrations to those who wish to utilize them for assays and further experimental studies.