Zhou Xuefeng, Xing Xuejiao, Hou Jingli, Liu Jianhua
School of Life Science & Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.
PLoS One. 2017 Jun 29;12(6):e0179842. doi: 10.1371/journal.pone.0179842. eCollection 2017.
While many data are available on genes encoding proteins for degradation of hydrocarbons in bacteria, the impact of alkane on transporter protein expression is unclear. Pseudomonas aeruginosa SJTD-1 is a strain that can consume medium- and long-chain n-alkanes. In order to study the proteins involved in n-octadecane uptake, we use iTRAQ and label free comparative proteomics analysis to identify the proteins of alkane uptake in response to n-octadecane (C18) comparing with n-hexadecane (C16) in P. aeruginosa SJTD-1. A total of 1102 and 1249 proteins were identified by iTRAQ-based and label free quantitative methodologies, respectively. By application of 1.5 (iTRAQ) or 2-fold (label free) for upregulated and 0.65 (iTRAQ) or 0.5-fold (label free) for downregulated cutoff values, 91 and 99 proteins were found to be differentially expressed comparing SJTD-1 cultivated on C18 with C16 respectively. There are six proteins with the common differential expression by iTRAQ and label free-based methods. Results of bioinformational analysis suggested the involvement of bacterial chemotaxis in responds to C18. Additionally, quantitative reverse transcriptase PCR (qRT-PCR) results confirmed C18-induced change in levels of FleQ, FliC, NirS, FadL and FadD proteins and the role of the proteins in n-octadecane uptake was further discussed in P. aeruginosa. In conclusion, results of the present study provided information about possible target-related proteins of bacterial chemotaxis, swimming performance, alkane transport to stimulus of n-ctadecane rather than n-hexadecane in P. aeruginosa SJTD-1.
虽然有许多关于编码细菌中烃降解蛋白的基因的数据,但烷烃对转运蛋白表达的影响尚不清楚。铜绿假单胞菌SJTD-1是一种能够消耗中链和长链正构烷烃的菌株。为了研究参与正十八烷摄取的蛋白质,我们使用iTRAQ和无标记比较蛋白质组学分析,以鉴定铜绿假单胞菌SJTD-1中响应正十八烷(C18)与正十六烷(C16)摄取的烷烃相关蛋白质。分别通过基于iTRAQ和无标记定量方法鉴定出1102和1249种蛋白质。通过将上调的截止值应用为1.5(iTRAQ)或2倍(无标记),下调的截止值应用为0.65(iTRAQ)或0.5倍(无标记),发现分别在C18和C16上培养的SJTD-1中有91和99种蛋白质差异表达。有六种蛋白质通过iTRAQ和基于无标记的方法具有共同的差异表达。生物信息分析结果表明细菌趋化性参与了对C18的反应。此外,定量逆转录聚合酶链反应(qRT-PCR)结果证实了C18诱导的FleQ、FliC、NirS、FadL和FadD蛋白水平的变化,并进一步讨论了这些蛋白在铜绿假单胞菌中对正十八烷摄取的作用。总之,本研究结果提供了关于铜绿假单胞菌SJTD-1中细菌趋化性、游动性能、烷烃转运以响应正十八烷而非正十六烷刺激的可能靶点相关蛋白的信息。