Time-resolved proteomic analysis of quorum sensing in Vibrio harveyi.
Time-resolved proteomic analysis of quorum sensing in Vibrio harveyi.
复制标题
DOI：
10.1039/c5sc03340c
复制
发表时间：
2016-03-01
期刊：
影响因子：
8.4
通讯作者：
Tirrell DA
中科院分区：
文献类型：
--
作者：
Bagert JD;van Kessel JC;Sweredoski MJ;Feng L;Hess S;Bassler BL;Tirrell DA
生物正交非规范氨基酸标记允许对哈维弧菌群体感应进行时间分辨蛋白质组学分析。细菌利用一种称为群体感应的化学交流过程来评估它们的种群密度，并根据群落中细胞数量和物种组成的波动改变它们的行为。在这项工作中，我们通过生物正交非规范氨基酸标记（BONCAT）鉴定了模式生物Vibrio harveyi群体感应调节的蛋白质组。BONCAT可以测量蛋白质组动力学，时间分辨率为几分钟。我们使用BONCAT来描述哈维氏菌从个体行为到群体行为的时间依赖性转变。我们确定了176个群体感应调节蛋白在早期、中期和晚期的过渡阶段，我们绘制了群体感应蛋白在转录和转录后机制控制下的时间变化。对鉴定的蛋白进行分析，发现86种已知和90种新的群体感应调节蛋白具有不同的功能，包括转录因子、趋化蛋白、运输蛋白和参与铁稳态的蛋白。
Bio-orthogonal non-canonical amino acid tagging allows time-resolved proteomic analysis of quorum sensing in Vibrio harveyi. Bacteria use a process of chemical communication called quorum sensing to assess their population density and to change their behavior in response to fluctuations in the cell number and species composition of the community. In this work, we identified the quorum-sensing-regulated proteome in the model organism Vibrio harveyi by bio-orthogonal non-canonical amino acid tagging (BONCAT). BONCAT enables measurement of proteome dynamics with temporal resolution on the order of minutes. We deployed BONCAT to characterize the time-dependent transition of V. harveyi from individual- to group-behaviors. We identified 176 quorum-sensing-regulated proteins at early, intermediate, and late stages of the transition, and we mapped the temporal changes in quorum-sensing proteins controlled by both transcriptional and post-transcriptional mechanisms. Analysis of the identified proteins revealed 86 known and 90 new quorum-sensing-regulated proteins with diverse functions, including transcription factors, chemotaxis proteins, transport proteins, and proteins involved in iron homeostasis.