幽门螺杆菌适应性定植蒙古沙鼠前、后的蛋白质组学研究

Comparative proteomics analysis of Helicobacter pylori after adaptive colonization in Mongolian gerbils

摘要:

为比较研究幽门螺杆菌(Hp)适应性定植蒙古沙鼠前后的蛋白表达谱变化,将Hp临床分离株感染沙鼠,并体内连续传代,驯化一株高适应性菌株,然后采用双向电泳和质谱技术对适应性变化前后两株菌的差异蛋白进行鉴定。结果表明,随着Hp临床株在沙鼠体内的连续传代,感染率逐渐升高,第10代后,感染率稳定在90%以上。适应性定植后,Hp蛋白谱发生了变化。在选择的5个候选差异蛋白点中,共鉴定出4个蛋白,分别为Hp菌的功能未知的HP0318编码蛋白、氢化酶表达/形成蛋白(hypB)、异柠檬酸脱氢酶(icd)、ADP-L-D-甘露庚糖表异构酶(rfaD)。上述鉴定蛋白可能与Hp适应性定植具有很大的相关性。

Abstract:

To study adaptive protein variation of H. pylori after colonization in Mongolian gerbils, Firstly, Clinical isolated strain M0 of H. pylori were inoculated into Mongolian gerbils and acclimated through serial passages in vivo for procuring an adaptive colonization H. pylori strain. Then, two-dimensional electrophoresis (2-DE) and mass spectrometry(MS) was taken to separate and identify the global proteins significantly changed between H. pylori strain M0and the adaptive strain. Through serial passages the infectious rate increased from about 2/10 to 9/10 and a adaptive colonization strain M₁₃ has been abtained. Comparative proteomic technology display that the proteinogram of H. pylori have changed after colonization in gerbils. Out of 5 differential protein spots cut out of gels for MALDI-TOF-MS identification, 4 spots were successfully identified, among which, Icd, RfaD and HP0318 were significantly higher in M₁₃ compared with M₀, while only HypB were found in M₁₃. So far, HP0318 is a conserved hypothetical protein. These proteins may be important factors of H. pylori for adaptive colonization.