Quantitative Proteomics Reveals the Temperature-Dependent Proteins Encoded by a Series of Cluster Genes in Thermoanaerobacter Tengcongensis
Quantitative Proteomics Reveals the Temperature-Dependent Proteins Encoded by a Series of Cluster Genes in Thermoanaerobacter Tengcongensis
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DOI：
10.1074/mcp.m112.025817
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发表时间：
2013-08-01
影响因子：
7
通讯作者：
Liu, Siqi
中科院分区：
文献类型：
--
作者：
Chen, Zhen;Wen, Bo;Liu, Siqi
嗜热菌蛋白质组的全面定量信息是了解其高温生存机制的重要来源。腾冲嗜热厌氧菌（Thermoanaerobacteriumtengcongensis，T. tengcongensis）的蛋白质丰度变化进行了定量研究。通过优化用于相对和绝对定量蛋白质组学（iTRAQ）的同量异位素标记程序，如高pH反相高效液相色谱（HPLC）肽段分级分离、LTQ Orbitrap Velos MS串联质谱采集模式以及定量算法的评价，获得了高质量的肽段定量信息。总共鉴定了1589种独特的蛋白质，并将251种确定为温度依赖性蛋白质。对这些温度依赖蛋白的对应基因的基因组定位分析表明，超过30%的温度依赖蛋白是具有相关生物学功能的连续单元，它们可能形成T.腾冲RNA测序（RNA-seq）结果进一步证明这些簇基因是共转录的，其mRNA丰度随温度的变化趋势与蛋白质组学结果相似，表明温度依赖性蛋白与相应的转录状态高度相关。因此，操纵子调控可能是T.腾冲生存。此外，对差异蛋白质组功能的评价表明，随着温度的升高，质膜上参与硫呼吸的蛋白质丰度降低，糖酵解相关蛋白质丰度增加。T.因此推测高温下腾冲藻的呼吸链反应不是主要的。
Comprehensive and quantitative information of the thermophile proteome is an important source for understanding of the survival mechanism under high growth temperature. Thermoanaerobacter tengcongensis (T. tengcongensis), a typical anaerobic thermophilic eubacterium, was selected to quantitatively evaluate its protein abundance changes in response to four different temperatures. With optimized procedures of isobaric tags for relative and absolute quantitation quantitative proteomics (iTRAQ), such as peptide fractionation with high-pH reverse phase (RP) high performance liquid chromatography (HPLC), tandem MS acquisition mode in LTQ Orbitrap Velos MS, and evaluation of the quantification algorithms, high quality of the quantitative information of the peptides identified were acquired. In total, 1589 unique proteins were identified and defined 251 as the temperature-dependent proteins. Analysis of genomic locations toward the correspondent genes of these temperature-dependent proteins revealed that more than 30% were contiguous units with relevant biological functions, which are likely to form the operon structures in T. tengcongensis. The RNA sequencing (RNA-seq) data further demonstrated that these cluster genes were cotranscribed, and their mRNA abundance changes responding to temperature exhibited the similar trends as the proteomic results, suggesting that the temperature-dependent proteins are highly associated with the correspondent transcription status. Hence, the operon regulation is likely an energy-efficient mode for T. tengcongensis survival. In addition, evaluation to the functions of differential proteomes indicated that the abundance of the proteins participating in sulfur-respiration on the plasma membrane was decreased as the temperature increased, whereas the glycolysis-related protein abundance was increased. The energy supply in T. tengcongensis at high temperature is, therefore, speculated not mainly through the respiration chain reactions.