A Biochemical Lanthanide-Encoding Approach Enables Quantitative Monitoring of the Bacterial Response to Vancomycin Treatment
A Biochemical Lanthanide-Encoding Approach Enables Quantitative Monitoring of the Bacterial Response to Vancomycin Treatment
复制标题
生化镧系元素编码方法能够定量监测细菌对万古霉素治疗的反应
DOI：
10.1021/acs.biochem.0c00614
复制
发表时间：
2020
期刊：
影响因子：
2.9
通讯作者：
Qiuquan Wang
中科院分区：
文献类型：
--
作者：
Weitong Zhao;Yong Liang;Xiaowen Yan;Limin Yang;Qiuquan Wang
病原菌不仅具有致病攻击的机制，而且还具有防御外源入侵的机制，其中细菌细胞壁是攻击和防御的前线。我们开发了一种生化镧系元素编码方法来量化非规范氨基酸 (d-X)，该氨基酸以小比例编辑到细菌细胞壁中新生肽聚糖 UDP-MurNAc-五肽的末端酰基-d-Ala-d-X 中。这种方法克服了流行的光学成像和传统的基于高效液相色谱的方法所遇到的定量和准确性问题的困难。新合成的叠氮化物-d-Leu和酮-d-Met与炔基-d-Ala一起进行代谢组装，然后由相应制造的DBCO-DOTA-Gd、H2NO-DOTA-Eu和叠氮化物-DOTA-Sm标签进行生物正交编码。该方法允许使用 158Gd、153Eu 和 154Sm 物种非特异性同位素稀释电感耦合等离子体质谱法直接对细胞壁中的 d-X 进行原位定量，避免任何可能诱导 d-X 外消旋化的繁琐且复杂的“细胞破碎”预处理程序。获得的有关 d-X 的位点特异性和准确的原位信息能够定量监测金黄色葡萄球菌遇到万古霉素时的细菌反应，表明在确定结构和成分依赖性细菌抗生素耐药机制后，叠氮化物-d-Leu 和酮-d-Met 的组装量更为重要。此外，我们发现万古霉素和d-Ala联合使用可以恢复万古霉素的药效，可能是对抗万古霉素中度耐药的明智而简单的方法。金黄色葡萄球菌。
A pathogenic bacterium has its own mechanisms for not only pathogenic attack but also exogenous invasion defense, in which the bacterial cell wall is the front line of attack and defense. We developed a biochemical lanthanide-encoding approach to quantify the uncanonicald-amino acid (d-X) that was edited in a small proportion into the terminal acyl-d-Ala-d-X of nascent peptidoglycan UDP-MurNAc-pentapeptides in the bacterial cell wall. This approach overcomes the difficulties regarding quantification and accuracy issues encountered by the popular optical imaging and traditional high-performance liquid chromatography-based methods. Newly synthesizedazide-d-Leu andketone-d-Met were used together withalkynyl-d-Ala for their metabolic assembly and then bioorthogonally encoded by the correspondingly fabricatedDBCO-DOTA-Gd,H2NO-DOTA-Eu, andazide-DOTA-Sm tags. This approach allows direct quantification of thed-X in situ in the cell wall using158Gd,153Eu, and154Sm species-unspecific isotope dilution inductively coupled plasma mass spectrometry, avoiding any tedious and complex “cell-broken” pretreatment procedures that might induce racemization of thed-X. The obtained site-specific and accurate in situ information about thed-X enables quantitative monitoring of the bacterial response whenStaphylococcus aureusmeets vancomycin, showing that the amounts ofazide-d-Leu andketone-d-Met assembled are more important after determining the structure- and composition-dependent bacterial antibiotic resistance mechanisms. In addition, we found that the combined use of vancomycin andd-Ala restores the efficacy of vancomycin and might be a wise and simple way to combat vancomycin intermediate-resistantS. aureus.