黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析

罗文英; 刘建军; 丁瑜; 杨拉维; 任晓虎

doi:10.16016/j.1000-5404.201701023

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黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析

基础医学 | 更新时间：2026-01-09

- 黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析
- Comparative proteomics analysis between mucoid and non-mucoid Pseudomonas aeruginosa strains
- 第三军医大学学报 2017年39卷第13期页码：1345-1351
- 作者机构：
  
  1.524001 广东湛江，广东医科大学附属医院检验科
  2.528055 深圳，深圳市疾病预防控制中心毒理实验室
  3.524001 广东湛江，广东医科大学附属医院临床研究中心
- 作者简介：
  
  [ 任晓虎，E-mail: slim54@126.com REN Xiaohu, E-mail:slim54@126.com
- 基金信息：
  
  国家自然科学基金青年科学基金(81301478);广东省湛江市非资助科技计划项目(2015B01042);广东省医学科学技术研究基金项目(B2017055)
- DOI：10.16016/j.1000-5404.201701023
  中图分类号： R372;R378.991;R394.33
- 收稿：2017-01-05，
  
  修回：2017-03-08，
  
  纸质出版：2017-07-15
- 稿件说明：
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罗文英, 刘建军, 丁瑜, 等. 黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析[J]. 第三军医大学学报, 2017,39(13):1345-1351.

Wenying LUO, Jianjun LIU, Yu DING, et al. Comparative proteomics analysis between mucoid and non-mucoid Pseudomonas aeruginosa strains[J]. Journal of Third Military Medical University, 2017, 39(13): 1345-1351.
罗文英, 刘建军, 丁瑜, 等. 黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析[J]. 第三军医大学学报, 2017,39(13):1345-1351. DOI： 10.16016/j.1000-5404.201701023.

Wenying LUO, Jianjun LIU, Yu DING, et al. Comparative proteomics analysis between mucoid and non-mucoid Pseudomonas aeruginosa strains[J]. Journal of Third Military Medical University, 2017, 39(13): 1345-1351. DOI： 10.16016/j.1000-5404.201701023.

摘要

目的

探讨黏液型与非黏液型铜绿假单胞菌(

Pseudomonas aeruginosa

)的差异蛋白谱，以发现黏液型

潜在的生物标志物。

方法

采用来自于临床囊性纤维化患者分离的3株黏液型

菌株，脉冲场凝胶电泳以确定它们的同源性；采用同位素相对和绝对定量标记(isobaric tags for relative and absolute quantitation，iTRAQ)结合二维液相色谱串联质谱(two-dimensional liquid chromatography-tandem mass spectrometry，2D LC-MS/MS)技术分析3株黏液型与对照组1株非黏液型

的差异蛋白谱，并采用RT-qPCR方法对5个共同差异蛋白进行验证。

结果

3株黏液型

菌株不具有同源性，iTRAQ结果没有明显差异。3组共有219个上调和101个下调差异蛋白。pa2815、pa2018、dnak、pa5046和acef基因转录水平与iTRAQ结果一致。

结论

不同黏液表型

的iTRAQ结果没有明显差异，219个上调和101个下调差异蛋白可以作为研究黏液型与非黏液型

致病机制的候选生物标志物。

Abstract

Objective

To investigate the differences in protein profiling between mucoid and non-mucoid

Pseudomonas aeruginosa

(

)

in order to find the potential biological markers of mucoid

Methods

Three strains of mucoid

were isolated from the sputum of cystic fibrosis patients

and then pulsed field gel electrophoresis was used to determine their homology. Isobaric tags for relative and absolute quantitation (iTRAQ) labeling strategy combined with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) was used to investigate the differential protein abundances between the 3 mucous and 1 non-mucoid

strains. Differential expression of 5 common differential genes was verified by using RT-qPCR.

Results

There was no homology in the 3 mucoid

strains. No significant differences were found in the iTRAQ results of them. There were 219 up-regulated and 101 down-regulated proteins identified among the 3 strains. The transcriptional levels of pa2815

pa2018

dnak

pa5046 and acef genes were consistent with the results of iTRAQ.

Conclusion

There are no significant differences in the iTRAQ results of different mucous

strains. The obtained 219 up-regulated and 101 down-regulated differentially expressed proteins can be used as candidate biomarkers for the study of the pathogenesis of mucoid and n

on-mucoid

关键词

Keywords

references

DRISCOLL J A, BRODY S L, KOLLEF M H. The epidemiology, pathogenesis and treatment of Pseudomonas aeruginosa infections[J ] . Drugs, 2007, 67(3): 351-368. DOI:10.2165/00003495-200767030-00003.

BJARNSHOLT T, JENSEN P Ø, FIANDACA M J, et al. Pseudomonas aeruginosa biofilms in the respiratory tract of cystic fibrosis patients[J ] . Pediatr Pulmonol, 2009, 44(6): 547-558. DOI:10. 1002/ppul.21011.

MA L, CONOVER M, LU H, et al. Assembly and development of the Pseudomonas aeruginosa biofilm matrix[J ] . PLoS Pathog, 2009, 5(3): e1000354. DOI:10.1371/journal.ppat.1000354.

MA L, WANG J, WANG S, et al. Synthesis of multiple Pseudomonas aeruginosa biofilm matrix exopolysaccharides is post-transcriptionally regulated[J ] . Environ Microbiol, 2012, 14(8): 1995-2005. DOI:10.1111/j.1462-2920.2012.02753.x.

MANN E E, WOZNIAK D J. Pseudomonas biofilm matrix composition and niche biology[J]. FEMS Microbiol Rev, 2012, 36(4): 893-916. DOI:10.1111/j.1574-6976.2011.00322.x.

SONG Z, WU H, CIOFU O, et al. Pseudomonas aeruginosa alginate is refractory to Th1 immune response and impedes host immune clearance in a mouse model of acute lung infection[J ] . J Med Microbiol, 2003, 52(Pt 9): 731-740. DOI:10.1099/jmm.0.05122-0.

GOODMAN A L, LORY S. Analysis of regulatory networks in Pseudomonas aeruginosa by genomewide transcriptional profiling[J ] . Curr Opin Microbiol, 2004, 7(1): 39-44. DOI:10.1016/j.mib.2003.12.009.

HANNA S L, SHERMAN N E, KINTER M T, et al. Comparison of proteins expressed by Pseudomonas aeruginosa strains representing initial and chronic isolates from a cystic fibrosis patient: an analysis by 2-D gel electrophoresis and capillary column liquid chromatography-tandem mass spectrometry[J ] . Microbiology (Reading, Engl), 2000, 146(Pt 10): 2495-2508. DOI:10.1099/00221287-146-10-2495.

WANG W S, LIU X H, LIU L X, et al. iTRAQ-based quantitative proteomics reveals myoferlin as a novel prognostic predictor in pancreatic adenocarcinoma[J]. J Proteomics, 2013, 91: 453-465. DOI:10.1016/j.jprot.2013.06.032.

PFAFFL M W, HORGAN G W, DEMPFLE L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR[J]. Nucleic Acids Res, 2002, 30(9): e36. DOI:10.1093/nar/30.9.e36.

LIU H, SUN W B, LIANG R B, et al. iTRAQ-based quantitative proteomic analysis of Pseudomonas aeruginosa SJTD-1: A global response to n-octadecane induced stress[J ] . J Proteomics, 2015, 123: 14-28. DOI:10.1016/j.jprot.2015.03. 034.

WU W W, WANG G, BAEK S J, et al. Comparative study of three proteomic quantitative methods, DIGE, cICAT, and iTRAQ, using 2D gel-or LC-MALDI TOF/TOF[J]. J Proteome Res, 2006, 5(3): 651-658. DOI:10.1021/pr050405o.

MORITA Y, KOMORI Y, MIMA T, et al. Construction of a series of mutants lacking all of the four major mex operons for multidrug efflux pumps or possessing each one of the operons from Pseudomonas aeruginosa PAO1: MexCD-OprJ is an inducible pump[J ] . FEMS Microbiol Lett, 2001, 202(1): 139-143. DOI:10.1111/j.1574-6968.2001.tb10794.x.

BATTAILE K P, NGUYEN T V, VOCKLEY J, et al. Structures of isobutyryl-CoA dehydrogenase and enzyme-product complex: comparison with isovaleryl-and short-chain acyl-CoA dehydrogenases[J]. J Biol Chem, 2004, 279(16): 16526-16534. DOI:10.1074/jbc.M400034200.

SWIGONOVÁ Z, MOHSEN A W, VOCKLEY J. Acyl-CoA dehydrogenases: Dynamic history of protein family evolution[J]. J Mol Evol, 2009, 69(2): 176-193. DOI:10.1007/s00239-009-9263-0.

ZHANG J, ZHANG W, ZOU D, et al. Cloning and functional characterization of ACAD-9, a novel member of human acyl-CoA dehydrogenase family[J]. Biochem Biophys Res Commun, 2002, 297(4): 1033-1042. DOI:10.1016/s0006-291x(02)02336-7.

VALENCIANO S, LUCAS J R, PEDREGOSA A, et al. Induction of beta-oxidation enzymes and microbody proliferation in Aspergillus nidulans[J]. Arch Microbiol, 1996, 166(5): 336-341. DOI:10.1007/s002030050392.

FERNANDO D M, KUMAR A. Resistance-nodulation-division multidrug efflux pumps in gram-negative bacteria: role in virulence[J]. Antibiotics (Basel). 2013, 2:163-181. DOI: 10.3390/antibiotics2010163..

VAN BAMBEKE F, BALZI E, TULKENS P M. Antibiotic efflux pumps[J]. Biochem Pharmacol, 2000, 60(4): 457-470. DOI:10.1016/s0006-2952(00)00291-4.

WOLTER D J, SMITH-MOLAND E, GOERING R V, et al. Multidrug resistance associated with mexXY expression in clinical isolates of Pseudomonas aeruginosa from a Texas hospital[J ] . Diagn Microbiol Infect Dis, 2004, 50(1): 43-50. DOI:10.1016/j.diagmicrobio.2004.05.004.

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