Global analysis of protein N-myristoylation and exploration of N-myristoyltransferase as a drug target in the neglected human pathogen Leishmania donovani.
Global analysis of protein N-myristoylation and exploration of N-myristoyltransferase as a drug target in the neglected human pathogen Leishmania donovani.
复制标题
DOI：
10.1016/j.chembiol.2015.01.003
复制
发表时间：
2015-03-19
影响因子：
--
通讯作者：
Tate EW
中科院分区：
文献类型：
--
作者：
Wright MH;Paape D;Storck EM;Serwa RA;Smith DF;Tate EW
N-肉豆蔻酰基转移酶（N - myristoyltransferase， NMT）通过脂质肉豆蔻酸酯附着在靶蛋白的N端来调节蛋白功能，是真核寄生虫如利什曼原虫的一个有前途的药物靶点。只有少数NMT底物在利什曼原虫中被表征，并且缺乏n -肉豆肉酰化的全球图景。在这里，我们在活寄生虫中使用炔功能化的肉豆汁酸模拟物进行代谢标记，然后进行下游点击化学和分析，以鉴定promastigote（细胞外）和amastigote（细胞内）生命阶段的脂化蛋白。定量化学蛋白质组学用于分析NMT抑制剂的靶标参与，并定义n -肉豆蔻酰基化蛋白的补体。我们的研究结果为NMT调控的多种途径和NMT抑制的多效性提供了新的见解。这项工作构成了利什曼原虫蛋白脂化的第一个全球实验分析，并揭示了nmt相关生物学的程度，尚未探索这种被忽视的人类病原体。通过化学敲低和定量蛋白质组学定义的n -肉豆酸盐化蛋白质组学证明NMT是利什曼原虫的药物靶点。Wright等人使用代谢标记与炔烃-肉豆酸盐类似物、点击化学和蛋白质组学来鉴定利什曼原虫两个生命阶段的脂肪化蛋白。n -肉豆蔻酰基转移酶抑制的定量分析和脂化位点的鉴定确定了这种人类病原体的n -肉豆蔻酰基化蛋白质组。
N-Myristoyltransferase (NMT) modulates protein function through the attachment of the lipid myristate to the N terminus of target proteins, and is a promising drug target in eukaryotic parasites such as Leishmania donovani. Only a small number of NMT substrates have been characterized in Leishmania, and a global picture of N-myristoylation is lacking. Here, we use metabolic tagging with an alkyne-functionalized myristic acid mimetic in live parasites followed by downstream click chemistry and analysis to identify lipidated proteins in both the promastigote (extracellular) and amastigote (intracellular) life stages. Quantitative chemical proteomics is used to profile target engagement by NMT inhibitors, and to define the complement of N-myristoylated proteins. Our results provide new insight into the multiple pathways modulated by NMT and the pleiotropic effects of NMT inhibition. This work constitutes the first global experimental analysis of protein lipidation in Leishmania, and reveals the extent of NMT-related biology yet to be explored for this neglected human pathogen. Alkyne-tagged probes for protein lipidation are applied in Leishmania donovani A global profile of protein lipidation in two life stages is presented N-Myristoylated proteome defined by chemical knockdown and quantitative proteomics Evidence for NMT as a drug target in the leishmaniases Wright et al. use metabolic tagging with an alkyne-myristate analog, click chemistry, and proteomics to identify lipidated proteins in two life stages of the parasite Leishmania donovani. Quantitative profiling of N-myristoyltransferase inhibition and identification of lipidation sites define the N-myristoylated proteome in this human pathogen.
登录
查看更多内容
影响因子：
7.3
作者：
Brand, Stephen;Cleghorn, Laura A. T.;McElroy, Stuart P.;Robinson, David A.;Smith, Victoria C.;Hallyburton, Irene;Harrison, Justin R.;Norcross, Neil R.;Spinks, Daniel;Bayliss, Tracy;Norval, Suzanne;Stojanovski, Laste;Torrie, Leah S.;Frearson, Julie A.;Brenk, Ruth;Fairlamb, Alan H.;Ferguson, Michael A. J.;Read, Kevin D.;Wyatt, Paul G.;Gilbert, Ian H.
通讯作者：
Gilbert, Ian H.
影响因子：
4.9
作者：
Heal, William P.;Jovanovic, Biljana;Tate, Edward W.
通讯作者：
Tate, Edward W.
影响因子：
3.8
作者：
Galvin, Brendan D.;Li, Zhiru;Carlow, Clotilde K. S.
通讯作者：
Carlow, Clotilde K. S.
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
14.8
作者：
Heal, William P.;Wright, Megan H.;Tate, Edward W.
通讯作者：
Tate, Edward W.