Analysis of the protective capacity of three Streptococcus suis proteins induced under divalent-cation-limited conditions
Analysis of the protective capacity of three Streptococcus suis proteins induced under divalent-cation-limited conditions
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DOI：
10.1128/iai.00987-07
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发表时间：
2008-04-01
影响因子：
3.1
通讯作者：
Barbe, Jordi
中科院分区：
文献类型：
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作者：
Aranda, Jesus;Garrido, Maria Elena;Barbe, Jordi
猪链球菌是一种革兰氏阳性病原体，可引起猪和人类的严重疾病。毒力S.基于从该生物体基因组的草图注释文件获得的信息，分离了编码推定的二价阳离子结合脂蛋白的SUIS 89/1591菌株。这些基因的产物，这是诱导二价阳离子剥夺，随后纯化，并分析其免疫原性和保护能力。发现所有三种蛋白质（SsuiDRAFT 0103、SsuiDRAFT 0174和SsuiDRAFT 1237）都具有免疫原性，但其中只有一种（SsuiDRAFT 0103）诱导针对相同S.猪种。此外，S.本研究克隆了猪链球菌（suissuiDRAFT 1240）基因（adcR），并纯化了其蛋白产物。用纯化的S. suis AdcR蛋白首次通过实验表明AdcR DNA结合序列对应于TTAACNRGTTAA基序。此外，证明了需要Zn 2+或Mn 2+而不是Fe来建立AdcR与其靶序列的体外结合，以及AdcR结合ssuiDRAFT 0103和ssuiDRAFT 12.57基因启动子而不是ssuiDRAFT 0174基因启动子的能力。综上所述，这些数据表明SsuiDRAFT 0103是针对S. SUIS和支持的初步结果表明，参与摄取二价阳离子而不是铁的细菌包膜蛋白可用于保护目的。
Streptococcus suis is a gram-positive pathogen that causes serious diseases in pigs and, in some cases, humans. Three genes of the virulent S. suis 89/1591 strain, encoding putative divalent-cation-binding lipoproteins, were isolated based on information obtained from the draft annotation files of this organism's genome. The products of these genes, which are inducible by divalent-cation deprivation, were subsequently purified, and their immunogenic and protective abilities were analyzed. All three proteins (SsuiDRAFT 0103, SsuiDRAFT 0174, and SsuiDRAFT 1237) were found to be immunogenic, but only one of them (SsuiDRAFT 0103) induced a significant protective response (87.5%, P = 0.01) against the same S. suis strain. Furthermore, the S. suis ssuiDRAFT 1240 gene (adcR), which encodes a predicted regulator of Zn2+ and/or Mn2+ uptake in streptococci, was cloned, and its protein product was purified. Electrophoretic mobility shift assays with purified S. suis AdcR protein showed experimentally, for the first time, that the AdcR DNA-binding sequence corresponds to the TTAACNRGTTAA motif. In addition, a requirement for either Zn2+ or Mn2+, but not Fe, to establish in vitro binding of AdcR to its target sequence and the ability of AdcR to bind the ssuiDRAFT 0103 and ssuiDRAFT 12.57 gene promoters but not the promoter of the ssuiDRAFT 0174 gene were demonstrated. Taken together, these data suggest that SsuiDRAFT 0103 is a good candidate for vaccines against S. suis and support preliminary results indicating that bacterial envelope proteins involved in the uptake of divalent cations other than iron may be useful for protective purposes.