Protein intrinsic disorder in the acetylome of intracellular and extracellular Toxoplasma gondii.
Protein intrinsic disorder in the acetylome of intracellular and extracellular Toxoplasma gondii.
复制标题
DOI：
10.1039/c3mb25517d
复制
发表时间：
2013-04-05
影响因子：
--
通讯作者：
Uversky VN
中科院分区：
文献类型：
--
作者：
Xue B;Jeffers V;Sullivan WJ;Uversky VN
弓形虫是顶复门的专性细胞内寄生虫，该门包括许多具有医学和兽医重要性的物种。赖氨酸脱乙酰酶 (KDAC) 抑制剂表现出有效的抗寄生虫活性，这表明干扰赖氨酸乙酰化途径有望成为未来的药物靶向。我们最近使用高分辨率 LC-MS/MS 来鉴定通过乙酰赖氨酸抗体免疫纯化富集的寄生虫肽，产生了弓形虫细胞内增殖阶段的乙酰基组。在这项研究中，我们使用类似的方法通过鉴定非复制细胞外速殖子中的乙酰化蛋白来极大地扩展弓形虫乙酰组。细胞外寄生虫中乙酰化蛋白质的功能分解与细胞内寄生虫相似，涉及代谢、翻译和染色质生物学的蛋白质富集。现在，我们总共在多个亚细胞区室中检测到了多种具有不同功能的寄生虫蛋白上的 700 多个乙酰化位点。我们发现 96 种蛋白质在细胞内寄生虫中独特乙酰化，216 种蛋白质在细胞外寄生虫中独特乙酰化，177 种蛋白质在两种状态下都乙酰化。我们的研究结果表明，当速殖子从细胞内环境过渡到细胞外环境时，蛋白质组水平发生了巨大的变化，类似于记录这一过渡期间基因表达显着变化的报告。扩展的数据集还允许对针对这种翻译后修饰的赖氨酸残基周围的蛋白质内在紊乱程度进行彻底分析。这些分析表明，来自细胞外和细胞内速殖子的蛋白质中的乙酰化赖氨酸大部分位于相似的局部环境中，并且赖氨酸乙酰化优先发生在本质上无序或灵活的区域中。
Toxoplasma gondii is an obligate intracellular parasite of the phylum Apicomplexa, which includes a number of species of medical and veterinary importance. Inhibitors of lysine deacetylases (KDACs) exhibit potent antiparasitic activity, suggesting that interference with lysine acetylation pathways hold promise for future drug targeting. Using high resolution LC-MS/MS to identify parasite peptides enriched by immunopurification with acetyl-lysine antibody, we recently produced an acetylome of the proliferative intracellular stage of Toxoplasma. In this study, we used similar approaches to greatly expand the Toxoplasma acetylome by identifying acetylated proteins in non-replicating extracellular tachyzoites. The functional breakdown of acetylated proteins in extracellular parasites is similar to intracellular parasites, with an enrichment of proteins involved in metabolism, translation, and chromatin biology. Altogether, we have now detected over 700 acetylation sites on a wide variety of parasite proteins of diverse function in multiple subcellular compartments. We found 96 proteins uniquely acetylated in intracellular parasites, 216 uniquely acetylated in extracellular parasites, and 177 proteins acetylated in both states. Our findings suggest that dramatic changes occur at the proteomic level as tachyzoites transition from the intracellular to extracellular environment, similar to reports documenting significant changes in gene expression during this transition. The expanded dataset also allowed a thorough analysis of the degree of protein intrinsic disorder surrounding lysine residues targeted for this post-translational modification. These analyses indicate that acetylated lysines in proteins from extracellular and intracellular tachyzoites are largely located within similar local environments, and that lysine acetylation preferentially occurs in intrinsically disordered or flexible regions.
登录
查看更多内容
影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
6.8
作者：
Dunker, A. Keith;Silman, Israel;Sussman, Joel L.
通讯作者：
Sussman, Joel L.
影响因子：
4.4
作者：
Ahmad, Atta;Burns, Colin S.;Uversky, Vladimir N.
通讯作者：
Uversky, Vladimir N.
影响因子：
2.9
作者：
Dunker, AK;Lawson, JD;Obradovic, Z
通讯作者：
Obradovic, Z
影响因子：
7.4
作者：
Finkemeier, Iris;Laxa, Miriam;Sweetlove, Lee J.
通讯作者：
Sweetlove, Lee J.