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The Aeromonas salmonicida subsp. salmonicida exoproteome: global analysis, moonlighting proteins and putative antigens for vaccination against furunculosis

机译:鲑气单胞菌亚种。沙门氏菌外蛋白质组:整体分析,月光蛋白和推定的抗原用于预防糠fur病

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摘要

BACKGROUNDudAeromonas salmonicida subsp. salmonicida, the etiologic agent of furunculosis, is a major pathogen of fisheries worldwide. Despite the identification of several virulence factors the pathogenesis is still poorly understood. We have used high-throughput proteomics to display the differences between in vitro secretome of A. salmonicida wild-type (wt, hypervirulent, JF5054) and T3SS-deficient (isogenic ΔascV, extremely low-virulent, JF2747) strains in exponential (GP) and stationary (SP) phases of growth.ududRESULTSudAmong the different experimental conditions we obtained semi-quantitative values for a total of 2136 A. salmonicida proteins. Proteins of specific A. salmonicida species were proportionally less detected than proteins common to the Aeromonas genus or those shared with other Aeromonas species, suggesting that in vitro growth did not induce the expression of these genes. Four detected proteins which are unidentified in the genome of reference strains of A. salmonicida were homologous to components of the conjugative T4SS of A. hydrophila pRA1 plasmid. Polypeptides of three proteins which are specific to the 01-B526 strain were also discovered. In supernatants (SNs), the number of detected proteins was higher in SP (326 for wt vs 329 for mutant) than in GP (275 for wt vs 263 for mutant). In pellets, the number of identified proteins (a total of 1536) was approximately the same between GP and SP. Numerous highly conserved cytoplasmic proteins were present in A. salmonicida SNs (mainly EF-Tu, EF-G, EF-P, EF-Ts, TypA, AlaS, ribosomal proteins, HtpG, DnaK, peptidyl-prolyl cis-trans isomerases, GAPDH, Enolase, FbaA, TpiA, Pgk, TktA, AckA, AcnB, Mdh, AhpC, Tpx, SodB and PNPase), and several evidences support the theory that their extracellular localization was not the result of cell lysis. According to the Cluster of Orthologous Groups classification, 29% of excreted proteins in A. salmonicida SNs were currently poorly characterized.ududCONCLUSIONSudIn this part of our work we elucidated the whole in vitro exoproteome of hypervirulent A. salmonicida subsp. salmonicida and showed the secretion of several highly conserved cytoplasmic proteins with putative moonlighting functions and roles in virulence. All together, our results offer new information about the pathogenesis of furunculosis and point out potential candidates for vaccine development.
机译:背景 udAeromonas salicicida亚种。沙门氏菌是糠un病的病原体,是全世界渔业的主要病原体。尽管已鉴定出几种毒力因子,但其发病机理仍知之甚少。我们已使用高通量蛋白质组学来显示指数(GP)的沙门氏菌野生型(wt,高毒力,JF5054)和T3SS缺陷型(等基因ΔascV,极低毒力,JF2747)菌株的体外分泌基因组之间的差异 ud udRESULTS ud在不同的实验条件下,我们获得了总共2136个鲑鱼曲霉蛋白的半定量值。与沙门氏菌属或其他沙门氏菌属共有的蛋白质相比,特定沙门氏菌属物种的蛋白质按比例检测的少,这表明体外生长不会诱导这些基因的表达。在沙门氏菌参考菌株的基因组中未鉴定的四个检测到的蛋白质与嗜水链球菌pRA1质粒的缀合T4SS的成分同源。还发现了对01-B526菌株特异的三种蛋白质的多肽。在上清液(SNs)中,SP中检测到的蛋白质数量(野生型为326,突变体为329)要比GP中的更高(野生型为275,突变体为263)。在颗粒中,GP和SP之间鉴定出的蛋白质数量(总计1536)大致相同。鲑鱼曲霉SN中存在许多高度保守的细胞质蛋白(主要是EF-Tu,EF-G,EF-P,EF-Ts,TypA,AlaS,核糖体蛋白,HtpG,DnaK,肽基脯氨酰顺反异构酶,GAPDH ,烯醇化酶,FbaA,TpiA,Pgk,TktA,AckA,AcnB,Mdh,AhpC,Tpx,SodB和PNPase),并且一些证据支持其细胞外定位不是细胞裂解的结果的理论。根据直系同源群分类,目前对沙门氏菌SN中29%的分泌蛋白表征不佳。 ud ud结论 ud在我们的这一部分工作中,我们阐明了高毒性沙门氏菌亚种的整个体外蛋白质组。并显示出几种高度保守的胞质蛋白的分泌,这些蛋白具有假定的月光照功能和在毒力中的作用。总之,我们的结果提供了有关糠un病发病机理的新信息,并指出了疫苗开发的潜在候选者。

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