Lysine crotonylation is a newly discovered protein post-translational modification and was reported to share transferases and de-acylases with lysine acetylation. The acetyltransferase p300 was reported also contain crotonyltransferase activity and class I histone deacetylases were demonstrated to be the major histone decrotonylases. However, the decortonylases for non-histone proteins are unclear. Moreover, due to the lack of high-quality pan-antibodies, large-scale analysis of crotonylome still remains challenge. In this work, we comprehensively studied lysine crotonylome on both histones and non-histone proteins upon SAHA treatment and dramatically identified 10,163 lysine crotonylation sites in A549 cells. This is the first identification of 10,000s of lysine crotonylation sites and also the largest lysine crotonylome dataset up to now. Moreover, a parallel reaction monitoring-based experiment was performed for validation, which presented highly consistent results with the SILAC experiments. By intensive bioinformatic analysis, it was found that lysine crotonylation participate in a wide range of biological functions and processes. More importantly, it was revealed that both the crotonylation and acetylation level of most core histones sites and a number of non-histone proteins as well as some known substrates of class IIa and IIb HDACs were up-regulated after SAHA treatment. These results suggest that SAHA may have decrotonylation inhibitory activities on both histones and non-histone proteins by inhibiting HDACs.

中文翻译：

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超深的赖氨酸巴豆基因组通过SAHA处理揭示了组蛋白和非组蛋白的巴豆酰化增强

赖氨酸巴豆酰化是新发现的蛋白质翻译后修饰，据报道与赖氨酸乙酰化共享转移酶和去酰化酶。据报道，乙酰基转移酶p300还具有巴豆酰基转移酶活性，并且I类组蛋白脱乙酰基酶被证明是主要的组蛋白脱巴豆酰基化酶。但是，尚不清楚非组蛋白的去甲酰化酶。而且，由于缺乏高质量的泛抗体，对巴豆组的大规模分析仍然是挑战。在这项工作中，我们对SAHA处理后的组蛋白和非组蛋白的赖氨酸巴豆基蛋白质组进行了全面研究，并在A549细胞中显着鉴定了10,163个赖氨酸巴豆基化位点。这是10,000个赖氨酸巴豆酰化位点的首次鉴定，也是迄今为止最大的赖氨酸巴豆酰化酶数据集。此外，进行了基于平行反应监测的实验以进行验证，该实验与SILAC实验呈现出高度一致的结果。通过深入的生物信息学分析，发现赖氨酸巴豆酰化参与了广泛的生物学功能和过程。更重要的是，揭示了在SAHA处理后，大多数核心组蛋白位点和许多非组蛋白蛋白以及IIa和IIb类HDAC的某些已知底物的巴豆酰化和乙酰化水平均被上调。这些结果表明，SAHA可能通过抑制HDAC而对组蛋白和非组蛋白具有去甲酰基化抑制活性。通过深入的生物信息学分析，发现赖氨酸巴豆酰化参与了广泛的生物学功能和过程。更重要的是，揭示了在SAHA处理后，大多数核心组蛋白位点和许多非组蛋白蛋白以及IIa和IIb类HDAC的某些已知底物的巴豆酰化和乙酰化水平均被上调。这些结果表明，SAHA可能通过抑制HDAC而对组蛋白和非组蛋白具有去甲酰基化抑制活性。通过深入的生物信息学分析，发现赖氨酸巴豆酰化参与了广泛的生物学功能和过程。更重要的是，揭示了在SAHA处理后，大多数核心组蛋白位点和许多非组蛋白蛋白以及IIa和IIb类HDAC的某些已知底物的巴豆酰化和乙酰化水平均被上调。这些结果表明，SAHA可能通过抑制HDAC而对组蛋白和非组蛋白具有去甲酰基化抑制活性。结果表明，SAHA处理后，大多数核心组蛋白位点和许多非组蛋白的巴豆酰化和乙酰化水平以及一些已知的IIa和IIb类HDAC底物均被上调。这些结果表明，SAHA可能通过抑制HDAC而对组蛋白和非组蛋白具有去甲酰基化抑制活性。结果表明，SAHA处理后，大多数核心组蛋白位点和许多非组蛋白的巴豆酰化和乙酰化水平以及一些已知的IIa和IIb类HDAC底物均被上调。这些结果表明，SAHA可能通过抑制HDAC而对组蛋白和非组蛋白具有去甲酰基化抑制活性。