Structural, kinetic and proteomic characterization of acetyl phosphate-dependent bacterial protein acetylation.
Structural, kinetic and proteomic characterization of acetyl phosphate-dependent bacterial protein acetylation.
复制标题
DOI：
10.1371/journal.pone.0094816
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Wolfe AJ
中科院分区：
文献类型：
--
作者：
Kuhn ML;Zemaitaitis B;Hu LI;Sahu A;Sorensen D;Minasov G;Lima BP;Scholle M;Mrksich M;Anderson WF;Gibson BW;Schilling B;Wolfe AJ
真核生物中Nε-赖氨酸乙酰化的新观点是相对丰富的翻译后修饰（PTM），其对参与不同细胞过程的数千种蛋白质的功能、结构、稳定性和/或位置具有重大影响。该PTM通常被认为是通过将乙酰辅酶A（acCoA）的乙酰基给予赖氨酸残基的ε-氨基而产生的，所述赖氨酸残基的ε-氨基被赖氨酸乙酰转移酶和脱乙酰酶可逆地催化。在这里，我们提供了遗传，质谱，生化和结构的证据，Nε-赖氨酸乙酰化是一个同样丰富和重要的PTM在细菌中。应用最近开发的，无标记和全球质谱的方法，以同基因组的突变体，我们检测到乙酰化的数千个赖氨酸残基的数百个大肠杆菌蛋白质，参与不同的，往往是必不可少的细胞过程，包括翻译，转录和中央代谢。许多这些乙酰化调节乙酰磷酸（acP）依赖性的方式，最近报道的细菌Nε-赖氨酸乙酰化的机制提供了令人信服的证据。这些质谱数据，再加上晶体学，生物化学和其他质谱法的观察结果表明，这种ACP依赖性乙酰化是非酶促和特异性的，特异性由目标赖氨酸的可及性，反应性和三维微环境决定。晶体学证据表明，acP可以在活性位点和辅因子结合位点与蛋白质结合，但也可能在具有磷酸基部分的分子可以结合的任何地方结合。最后，我们提供的证据表明，ACP依赖的乙酰化可以影响关键酶的功能，包括甘油醛-3-磷酸脱氢酶，磷酸丙糖异构酶和RNA聚合酶。
The emerging view of Nε-lysine acetylation in eukaryotes is of a relatively abundant post-translational modification (PTM) that has a major impact on the function, structure, stability and/or location of thousands of proteins involved in diverse cellular processes. This PTM is typically considered to arise by the donation of the acetyl group from acetyl-coenzyme A (acCoA) to the ε-amino group of a lysine residue that is reversibly catalyzed by lysine acetyltransferases and deacetylases. Here, we provide genetic, mass spectrometric, biochemical and structural evidence that Nε-lysine acetylation is an equally abundant and important PTM in bacteria. Applying a recently developed, label-free and global mass spectrometric approach to an isogenic set of mutants, we detected acetylation of thousands of lysine residues on hundreds of Escherichia coli proteins that participate in diverse and often essential cellular processes, including translation, transcription and central metabolism. Many of these acetylations were regulated in an acetyl phosphate (acP)-dependent manner, providing compelling evidence for a recently reported mechanism of bacterial Nε-lysine acetylation. These mass spectrometric data, coupled with observations made by crystallography, biochemistry, and additional mass spectrometry showed that this acP-dependent acetylation is both non-enzymatic and specific, with specificity determined by the accessibility, reactivity and three-dimensional microenvironment of the target lysine. Crystallographic evidence shows acP can bind to proteins in active sites and cofactor binding sites, but also potentially anywhere molecules with a phosphate moiety could bind. Finally, we provide evidence that acP-dependent acetylation can impact the function of critical enzymes, including glyceraldehyde-3-phosphate dehydrogenase, triosephosphate isomerase, and RNA polymerase.
登录
查看更多内容
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1006/bbrc.2000.3000
发表时间：
2000-07-05
影响因子：
3.1
作者：
Frye, RA
通讯作者：
Frye, RA
DOI：
10.1111/j.1432-1033.1978.tb12074.x
发表时间：
1978-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
FOUCAULT, G;NAKANO, M;PUDLES, J
通讯作者：
PUDLES, J
影响因子：
15
作者：
DISABATO, G;JENCKS, WP
通讯作者：
JENCKS, WP
影响因子：
3.5
作者：
Da Re, SS;Deville-Bonne, D;Stock, JB
通讯作者：
Stock, JB