Quantitative and Structural Assessment of Histone Methyllysine Analogue Engagement by Cognate Binding Proteins Reveals Affinity Decrements Relative to Those of Native Counterparts.
Quantitative and Structural Assessment of Histone Methyllysine Analogue Engagement by Cognate Binding Proteins Reveals Affinity Decrements Relative to Those of Native Counterparts.
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同源结合蛋白对组蛋白甲基苷类似物参与的定量和结构评估揭示了相对于天然对应物的亲和力的降低。
DOI：
10.1021/acs.biochem.7b00926
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发表时间：
2018-01-23
期刊：
影响因子：
2.9
通讯作者：
Ruthenburg AJ
中科院分区：
文献类型：
--
作者：
Chen Z;Notti RQ;Ueberheide B;Ruthenburg AJ
甲基赖氨酸类似物 (MLA) 由工程半胱氨酸残基氨乙基化提供，是广泛使用的组蛋白甲基赖氨酸替代物，被认为是体外研究这些表观遗传标记的有效替代物。在这里，我们报告了三甲基赖氨酸 MLA 组蛋白与蛋白质结合伴侣复合物的第一个结构，量化了 MLA 与其天然甲基赖氨酸对应物之间的热力学差异，并证明这些差异可能会影响核小体水平结合的定性解释。尽管热力学基础不同，但使用两个甲基赖氨酸结合蛋白模块进行的定量测量表明，在这两种情况下，MLA 肽与相应的天然甲基赖氨酸物种相比都有显着的亲和力损失。 MLA 和甲基赖氨酸与 BPTF PHD 指（一种成熟的 H3K4me3 结合伴侣）复合时，采用不同的构象几何形状。在这种情况下，肽水平上约 13 倍的 Kd 差异转化为 MLA 类似物的核小体亲和力，超出了下拉格式的可检测范围，而通过天然化学连接安装的甲基赖氨酸种类则表现出强大的结合。因此，尽管它们的生产和商业可用性很容易，但当使用 MLA 代替天然甲基赖氨酸残基时，仍需要注意可能会改变结合亲和力。
Methyllysine analogs (MLAs), furnished by aminoethylation of engineered cysteine residues, are widely used surrogates of histone methyllysine and are considered to be effective proxies for studying these epigenetic marks in vitro. Here we report the first structure of a trimethyllysine MLA histone in complex with a protein binding partner, quantify the thermodynamic distinctions between MLAs and their native methyllysine counterparts, and demonstrate that these differences can compromise qualitative interpretations of binding at the nucleosome-level. Quantitative measurements with two methyllysine-binding protein modules reveal substantial affinity losses for the MLA peptides versus the corresponding native methyllysine species in both cases, although the thermodynamic underpinnings are distinct. MLA and methyllysine adopt distinct conformational geometries when in complex the BPTF PHD finger, a well-established H3K4me3 binding partner. In this case, a ~13-fold Kd difference at the peptide-level translates to nucleosomal affinities for MLA analogs that fall outside of the detectable range in a pull-down format, whereas the methyllysine species installed by native chemical ligation demonstrates robust binding. Thus, despite their facile production and commercial availability, there is a significant caveat of potentially altered binding affinity when MLAs are used in place of native methyllysine residues.
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影响因子：
3.9
作者：
van Nuland R;van Schaik FM;Simonis M;van Heesch S;Cuppen E;Boelens R;Timmers HM;van Ingen H
通讯作者：
van Ingen H
影响因子：
3.2
作者：
Chen, Zhonglei;Grzybowski, Adrian T.;Ruthenburg, Alexander J.
通讯作者：
Ruthenburg, Alexander J.
影响因子：
48
作者：
Nguyen, Uyen T. T.;Bittoval, Lenka;Mueller, Manuel M.;Fierz, Beat;David, Yael;Houck-Loomis, Brian;Feng, Vanessa;Dann, Geoffrey P.;Muir, Tom W.
通讯作者：
Muir, Tom W.
影响因子：
64.5
作者：
Ruthenburg AJ;Li H;Milne TA;Dewell S;McGinty RK;Yuen M;Ueberheide B;Dou Y;Muir TW;Patel DJ;Allis CD
通讯作者：
Allis CD
影响因子：
16.8
作者：
Musselman, Catherine A.;Lalonde, Marie-Eve;Cote, Jacques;Kutateladze, Tatiana G.
通讯作者：
Kutateladze, Tatiana G.