N-acetyl-L-methionine sulfoximine(1-)

• 分子结构分类: 有机化合物 - 有机酸及其衍生物 - 羧酸及其衍生物
• 英文名称: N-acetyl-L-methionine sulfoximine(1-)
• 英文别名: (2S)-2-acetamido-4-(methylsulfonimidoyl)butanoate
• 化学式: C7H13N2O4S-
• 分子量: 221.26
• InChiKey: NBUSDVOJMKDYAH-TZKMECQKSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  0.8
• 重原子数：
  14
• 可旋转键数：
  4
• 环数：
  0.0
• sp3杂化的碳原子比例：
  0.71
• 拓扑面积：
  119
• 氢给体数：
  2
• 氢受体数：
  5

反应信息

• 作为产物：
  描述：

  (2R)-N-[3-(2-acetylsulfanylethylimino)-3-oxidopropyl]-4-[[[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-4-hydroxy-3-phosphonatooxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]oxy-2-hydroxy-3,3-dimethylbutanimidate 、 L-蛋氨酸磺酸盐 生成 coenzyme A 、 氢(+1)阳离子 、 N-acetyl-L-methionine sulfoximine(1-)
  参考文献：
  名称：

  In Salmonella enterica, the Gcn5-Related Acetyltransferase MddA (Formerly YncA) Acetylates Methionine Sulfoximine and Methionine Sulfone, Blocking Their Toxic Effects

  摘要：

  蛋白质和小分子酰化反应在自然界中广泛存在。催化酰化反应的许多酶属于 Gcn5 相关 N-乙酰转移酶（GNAT；PF00583）家族，该家族以酵母 Gcn5 蛋白命名。肠道沙门氏菌 Typhimurium LT2 的基因组编码了 26 个 GNAT，其中 11 个没有已知的生理作用。在这里，我们提供了体内和体外证据，证明 MddA（methioninederivativedetoxifier；前身为 YncA）GNAT 在甲硫氨酸氧化形式（包括甲硫氨酸亚砜亚胺（MSX）和甲硫氨酸砜（MSO））的解毒过程中发挥作用。除非培养基中含有谷氨酰胺或蛋氨酸，否则 MSX 和 MSO 会抑制肠炎Δmdd 菌株的生长。我们使用玻璃分光光度法和质谱法证明 MddA 对 MSX 和 MSO 进行了乙酰化。AnmddA+ 菌株在 MSX 和谷氨酰胺存在下显示出双相生长动力学。在 ΔmddA 菌株中缺失两种氨基酸转运体（GlnHPQ 和 MetNIQ）后，在有 MSX 存在的情况下，该菌株恢复了生长。值得注意的是，MSO 是由 GlnHPQ 转运的，而不是由 MetNIQ 转运的。总之，MddA 是肠道病毒对氧化型蛋氨酸做出反应的机制，MddA 通过乙酰辅酶 A 依赖性乙酰化对氧化型蛋氨酸进行解毒。

  DOI：

  10.1128/jb.02311-14

文献信息

• Nontargeted in vitro metabolomics for high-throughput identification of novel enzymes in Escherichia coli
  作者：Daniel C Sévin、Tobias Fuhrer、Nicola Zamboni、Uwe Sauer

  DOI：10.1038/nmeth.4103

  日期：2017.2

  A method to screen proteins for enzymatic activity by incubating purified or overexpressed proteins with a metabolite extract and measuring changes in metabolite abundance using mass spectrometry enables high-throughput characterization of functionally uncharacterized proteins in Escherichia coli. Our understanding of metabolism is limited by a lack of knowledge about the functions of many enzymes. Here, we develop a high-throughput mass spectrometry approach to comprehensively profile proteins for in vitro enzymatic activity. Overexpressed or purified proteins are incubated in a supplemented metabolome extract containing hundreds of biologically relevant candidate substrates, and accumulating and depleting metabolites are determined by nontargeted mass spectrometry. By combining chemometrics and database approaches, we established an automated pipeline for unbiased annotation of the functions of novel enzymes. In screening all 1,275 functionally uncharacterized Escherichia coli proteins, we discovered 241 potential novel enzymes, 12 of which we experimentally validated. Our high-throughput in vitro metabolomics method is generally applicable to any purified protein or crude cell lysate of its overexpression host and enables performing up to 1,200 nontargeted enzyme assays per working day.

  通过将纯化或过表达的蛋白质与代谢物提取物孵育，并使用质谱法测量代谢物丰度的变化，筛选蛋白质的酶活性，这种方法能够对大肠杆菌中功能未定的蛋白质进行高通量表征。我们对代谢的理解受到许多酶功能缺乏了解的限制。在这里，我们开发了一种高通量质谱法，用于全面分析蛋白质的体外酶活性。将过表达或纯化的蛋白质与含有数百种生物相关候选底物的补充代谢物提取物孵育，并通过非靶向质谱法确定累积和耗竭的代谢物。通过结合化学计量学和数据库方法，我们建立了一个自动化的管道，用于对新型酶的功能进行无偏注解。在筛选所有1275种功能未定的大肠杆菌蛋白质时，我们发现了241种潜在的新型酶，其中12种通过实验验证。我们的高通量体外代谢组学方法通常适用于任何纯化的蛋白质或过表达宿主的粗细胞裂解液，每个工作日最多可以进行1200次非靶向酶检测。
• <scp>l</scp>-Methionine Sulfoximine, but Not Phosphinothricin, Is a Substrate for an Acetyltransferase (Gene PA4866) from <i>Pseudomonas aeruginosa</i>:  Structural and Functional Studies
  作者：Anna M. Davies、Renée Tata、Rebecca L. Beavil、Brian J. Sutton、Paul R. Brown

  DOI：10.1021/bi0615238

  日期：2007.2.1

  The gene PA4866 from Pseudomonas aeruginosa is documented in the Pseudomonas genome database as encoding a 172 amino acid hypothetical acetyltransferase. We and others have described the 3D structure of this protein (termed pita) [Davies et al. (2005) Proteins: Struct., Funct., Bioinf. 61, 677-679; Nocek et al., unpublished results], and structures have also been reported for homologues from Agrobacterium tumefaciens (Rajashankar et al., unpublished results) and Bacillus subtilis [Badger et al. (2005) Proteins: Struct., Funct., Bioinf. 60, 787-796]. Pita homologues are found in a large number of bacterial genomes, and while the majority of these have been assigned putative phosphinothricin acetyltransferase activity, their true function is unknown. In this paper we report that pita has no activity toward phosphinothricin. Instead, we demonstrate that pita acts as an acetyltransferase using the glutamate analogues l-methionine sulfoximine and l-methionine sulfone as substrates, with Km(app) values of 1.3 +/- 0.21 and 1.3 +/- 0.13 mM and kcat(app) values of 505 +/- 43 and 610 +/- 23 s-1 for l-methionine sulfoximine and l-methionine sulfone, respectively. A high-resolution (1.55 A) crystal structure of pita in complex with one of these substrates (l-methionine sulfoximine) has been solved, revealing the mode of its interaction with the enzyme. Comparison with the apoenzyme structure has also revealed how certain active site residues undergo a conformational change upon substrate binding. To investigate the role of pita in P. aeruginosa, a mutant strain, Depp4, in which pita was inactivated through an in-frame deletion, was constructed by allelic exchange. Growth of strain Depp4 in the absence of glutamine was inhibited by l-methionine sulfoximine, suggesting a role for pita in protecting glutamine synthetase from inhibition.