Investigating Peptidoglycan Recycling Pathways in Tannerella forsythia with N-Acetylmuramic Acid Bioorthogonal Probes.
Investigating Peptidoglycan Recycling Pathways in Tannerella forsythia with N-Acetylmuramic Acid Bioorthogonal Probes.
复制标题
DOI：
10.1021/acsinfecdis.2c00333
复制
发表时间：
2022-09-09
影响因子：
5.3
通讯作者：
Grimes, Catherine L.
中科院分区：
文献类型：
--
作者：
Wodzanowski, Kimberly A.;Hyland, Stephen N.;Chinthamani, Sreedevi;Sandles, Liam-Michael D.;Honma, Kiyonobu;Sharma, Ashu;Grimes, Catherine L.
关键词：
人类口腔微生物组是人类第二大微生物群落，含有700多种细菌，有助于消化并防止致病病原体的生长。其中一种口腔病原体，即连翘坦纳氏菌（Tannerella forsythia），与其他物种一起沿着导致牙周炎的发病机制。T.该菌不能产生其自身的N-乙酰胞壁酸（NAM）糖，这是肽聚糖生物合成所必需的，因此必须从与其共存的其他物种中除去NAM。在这里，我们探讨了T。通过将生物正交修饰的NAM摄取到其肽聚糖中，允许基于点击化学的细胞壁结构可视化。此外，我们还鉴定了T.与恶臭假单胞菌中发现的酶类似的酶。然后将这些同源物遗传转化到实验室安全的E.大肠杆菌菌株，导致非天然NAM类似物有效掺入肽聚糖骨架中，并单独或在人巨噬细胞存在下使其可视化。该菌株将有助于进一步研究T.以及其他共生细菌。T.该菌是在口腔微生物组中发现的构成“红色复合物”的病原体。在这篇论文中，我们能够用biothogronal点击化学标记这个物种，以荧光可视化细胞壁，证实预测的回收机制的存在。
The human oral microbiome is the second largest microbial community in humans, harboring over 700 bacterial species, which aid in digestion and protects from growth of disease-causing pathogens. One such oral pathogen, Tannerella forsythia, along with other species contribute to the pathogenesis of periodontitis. T. forsythia is unable to produce its own N-Acetylmuramic acid (NAM) sugar, essential for peptidoglycan biosynthesis, and therefore must scavenge NAM from other species it cohabitates with. Here, we explore the recycling potential of T. forsythia to uptake NAM with a bioorthogonal modification into its peptidoglycan, allowing for click chemistry-based visualization of the cell wall structure. Additionally, we identified NAM recycling enzyme homologs in T. forsythia similar to the enzymes found in Pseudomonas putida. These homologs were then genetically transformed into a laboratory safe E. coli strain, resulting in the efficient incorporation of unnatural NAM analogs into the peptidoglycan backbone and its visualization, alone or in the presence of human macrophages. This strain will be useful in further studies to probe NAM recycling and peptidoglycan scavenging pathways of T. forsythia and other cohabiting bacteria. T. forsythia is a pathogen found constituting the “red complex” in the oral microbiome. In this paper we are able to label this species with bioothogronal click chemistry to fluorescently visualize the cell wall, confirming the presence of predicted recycling machinery.
登录
查看更多内容
DOI：
10.1002/cpz1.80
发表时间：
2021-04
期刊：
Current protocols
影响因子：
--
作者：
Taylor JA;Santiago CC;Gray J;Wodzanowski KA;DeMeester KE;Biboy J;Vollmer W;Grimes CL;Salama NR
通讯作者：
Salama NR
影响因子：
3.7
作者：
Ruscitto A;Sharma A
通讯作者：
Sharma A
DOI：
10.1007/s13361-014-0880-5
发表时间：
2014-07-01
影响因子：
3.2
作者：
Knolhoff, Ann M.;Callahan, John H.;Croley, Timothy R.
通讯作者：
Croley, Timothy R.
影响因子：
3.2
作者：
Ruscitto, Angela;Hottmann, Isabel;Sharma, Ashu
通讯作者：
Sharma, Ashu
影响因子：
7.4
作者：
Clauser, KR;Baker, P;Burlingame, AL
通讯作者：
Burlingame, AL