Differences in specificity and selectivity between CBP and p300 acetylation of histone H3 and H3/H4.
Differences in specificity and selectivity between CBP and p300 acetylation of histone H3 and H3/H4.
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DOI：
10.1021/bi400684q
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发表时间：
2013-08-27
期刊：
影响因子：
2.9
通讯作者：
Andrews, Andrew J.
中科院分区：
文献类型：
--
作者：
Henry, Ryan A.;Kuo, Yin-Ming;Andrews, Andrew J.
虽然p300和CBP赖氨酸乙酰基转移酶经常被互换处理，一种酶无法补偿另一种酶的损失，这表明每种酶都有独特的作用。由于这些缺陷与组蛋白乙酰化的异常水平相一致，我们假设p300和CBP活性之间的关键差异是它们对组蛋白内赖氨酸的特异性/选择性的差异。利用无标记、基于定量质谱的技术，在我们预期在细胞中遇到的条件下（限制乙酰辅酶A或组蛋白），我们确定了H3和H4的每个赖氨酸处CBP和p300的动力学参数。我们的研究结果表明，虽然p300和CBP乙酰化H3和H4上的许多常见残基，但它们实际上具有非常不同的特异性，并且这些特异性取决于组蛋白或乙酰辅酶A是否是限制性的。用限制性H3进行的稳态实验表明，CBP和p300都能乙酰化H3 K14、H3 K18、H3 K23，其中p300的特异性比CBP高1010倍。利用四聚体作为底物，两种酶也乙酰化H4 K5、H4 K8、H4 K12和H4 K16。在限制性四聚体的情况下，CBP显示出更高的特异性，特别是在H3 K18，其中CBP特异性比p300高1032倍。在限制乙酰辅酶A的情况下，p300对H4 K16具有最高的特异性，其特异性比CBP高1018倍。CBP与p300介导的组蛋白赖氨酸残基乙酰化靶点的独特特异性的发现为理解其各自的生物学作用提供了新的模型，并可能为选择性治疗干预提供了机会。
Although p300 and CBP lysine acetyltransferases are often treated interchangeably, the inability of one enzyme to compensate for the loss of the other suggests unique roles for each. As these deficiencies coincide with aberrant levels of histone acetylation, we hypothesized that the key difference between p300 and CBP activity is differences in their specificity/selectivity for lysines within the histones. Utilizing a label-free, quantitative mass spectrometry based technique, we determined the kinetic parameters of both CBP and p300 at each lysine of H3 and H4, under conditions we would expect to encounter in the cell (either limiting acetyl-CoA or histone). Our results show that while p300 and CBP acetylate many common residues on H3 and H4, they do in fact possess very different specificities, and these specificities are dependent on whether histone or acetyl-CoA is limiting. Steady-state experiments with limiting H3 demonstrate that both CBP and p300 acetylate H3K14, H3K18, H3K23, with p300 having specificities up to 1010-fold higher than CBP. Utilizing tetramer as a substrate, both enzymes also acetylate H4K5, H4K8, H4K12, and H4K16. With limiting tetramer, CBP displays higher specificities, especially at H3K18, where CBP specificity is 1032-fold higher than p300. With limiting acetyl-CoA, p300 has the highest specificity at H4K16, where specificity is 1018-fold higher than CBP. This discovery of unique specificity for targets of CBP- vs p300-mediated acetylation of histone lysine residues presents a new model for understanding their respective biological roles and possibly an opportunity for selective therapeutic intervention.
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