An RNA Aptamer That Selectively Recognizes Symmetric Dimethylation of Arginine 8 in the Histone H3 N-Terminal Peptide
An RNA Aptamer That Selectively Recognizes Symmetric Dimethylation of Arginine 8 in the Histone H3 N-Terminal Peptide
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DOI：
10.1089/nat.2011.0300
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发表时间：
2011-06-01
影响因子：
4
通讯作者：
Yu, Jaehoon
中科院分区：
文献类型：
--
作者：
Hyun, Soonsil;Lee, Kyung Hyun;Yu, Jaehoon
N 端组蛋白尾部（尤其是组蛋白 H3）的表观遗传修饰对于染色质中靶基因的调控非常重要。检测组蛋白 H3 N 末端肽中这些修饰的具体方法是诊断和治疗目的的宝贵工具。作为抗体的替代品，RNA 适体对各种生物学相关靶标表现出兼容的结合亲和力和选择性。针对组蛋白 H3R8Me2sym 进行配体指数富集系统进化 (SELEX)。以生物素化形式制备模拟这种修饰组蛋白尾部的 14 个氨基酸肽，并进行 10 个 SELEX 选择循环。这产生了 4 个适体，其中一个（克隆 1）被发现对同源肽具有低纳摩尔结合亲和力（K-d = 12nM）。该适体的亲和力与针对不同修饰的组蛋白 H3 肽的 2 种市售抗体相当，并且它比抗体具有更高的选择性。
Epigenetic modifications of N-terminal histone tails, especially histone H3, are important for the regulation of the target genes in chromatin. Specific methods for detection of these modifications in histone H3 N-terminal peptides are valuable tools for diagnostic and therapeutic purposes. As an alternative to antibodies, RNA aptamers display compatible binding affinities and selectivites against various biologically relevant targets. Systematic evolution of ligands by exponential enrichment (SELEX) was performed against histone H3R8Me2sym. A 14-amino acid peptide that mimics this modified histone tail was prepared in a biotinylated form and 10 selection cycles of SELEX were carried out. This produced 4 aptamers, one of which (clone 1) was observed to have low nanomolar binding affinity (K-d = 12nM) against the cognate peptide. The affinity of this aptamer is comparable to 2 commercially available antibodies against differently modified histone H3 peptides and it displays a greater selectivity than the antibodies.