Hydrophobic cavity-directed azide-acetyllysine photochemistry for profiling non-histone interacting partners of bromodomain protein 1.
Hydrophobic cavity-directed azide-acetyllysine photochemistry for profiling non-histone interacting partners of bromodomain protein 1.
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疏水腔导向叠氮-乙酰赖氨酸光化学用于分析布罗莫结构域蛋白1的非组蛋白相互作用伴侣。
DOI：
10.1039/d2cb00043a
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发表时间：
2022-08-03
影响因子：
4.1
通讯作者：
Islam, Kabirul
中科院分区：
文献类型：
--
作者：
Kuwik, Jordan;Wagner, Shana;Sudhamalla, Babu;Debiec, Ronald;Islam, Kabirul
含溴结构域蛋白1（BRD 1）在染色质乙酰化、基因转录、红细胞生成和脑发育中起关键作用。BRD 1也与几种人类疾病有关，是癌症的治疗靶点。尽管已知溴结构域结合乙酰化组蛋白，但BRD 1的功能如何通过非组蛋白乙酰化调节尚未探索。为了鉴定BRD 1的非组蛋白乙酰组，我们通过琥珀抑制突变开发了携带光响应性4-叠氮基苯丙氨酸（AzF）的R585 AzF变体。我们证明了含AzF类似物的生物化学完整性及其交联人类细胞中存在的非组蛋白相互作用伙伴的能力。随后的蛋白质组学实验导致了代表不同信号通路的新型BRD 1相互作用组的鉴定。作为概念验证，我们验证了乙酰化PDIA 1蛋白作为BRD 1的真正结合伴侣。我们的工作表明，BRD 1与其他乙酰赖氨酸基序相互作用，超出了组蛋白中的特征。含溴结构域蛋白1（BRD 1）被改造为携带可光活化的氨基酸，用于表征非组蛋白相互作用伴侣。
Bromodomain containing protein 1 (BRD1) plays critical roles in chromatin acetylation, gene transcription, erythropoiesis, and brain development. BRD1 is also implicated in several human conditions and is a therapeutic target for cancer. Although, the bromodomain is known to bind acetylated histones, how the function of BRD1 is regulated via non-histone acetylation is unexplored. To identify the non-histone acetylome of BRD1, we develop an R585AzF variant carrying photo responsive 4-azido phenylalanine (AzF) via amber suppressor mutagenesis. We demonstrate biochemical integrity of the AzF-containing analogue and its ability to crosslink non-histone interacting partners present in human cells. Subsequent proteomic experiments led to the identification of the novel BRD1 interactome representing diverse signaling pathways. As a proof-of-concept demonstration, we validated acetylated PDIA1 protein as a bona fide binding partner of BRD1. Our work suggests that BRD1 interacts with additional acetyllysine motifs, beyond those characterized in histone proteins. Bromodomain-containing protein 1 (BRD1) was engineered to carry photo-activatable amino acid for characterization of non-histone interacting partners.
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影响因子：
5.6
作者：
Poplawski, Amanda;Hu, Kaifeng;Lee, Woonghee;Natesan, Senthil;Peng, Danni;Carlson, Samuel;Shi, Xiaobing;Balaz, Stefan;Markley, John L.;Glass, Karen C.
通讯作者：
Glass, Karen C.
影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
12.3
作者：
Fryland T;Christensen JH;Pallesen J;Mattheisen M;Palmfeldt J;Bak M;Grove J;Demontis D;Blechingberg J;Ooi HS;Nyegaard M;Hauberg ME;Tommerup N;Gregersen N;Mors O;Corydon TJ;Nielsen AL;Børglum AD
通讯作者：
Børglum AD
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.
影响因子：
15
作者：
Liu, Jun;Li, Shanshan;Wang, Lei
通讯作者：
Wang, Lei