幽门螺杆菌动物模型驯化菌株SS1与其初始菌株的差异蛋白质组分析

Comparative proteomic analysis of Helicobacter pylori animal model adapted SS1 and its original strain

目的:比较H pylori动物模型适应菌株SS1与其初发菌株10700间蛋白质组差异,试图获得H pylori在小鼠体内适应过程中可能与定植能力增强相关的蛋白特征.方法:在相同件下培养SS1菌株及其初始菌株,并制备全菌蛋白样品,相同实验条件下重复3次,样品经二维电泳分离,扫描获得数字化的电泳胶图像并用ImageMaster2D图像分析软件比较分析,识别SS1与初始菌株间的差异蛋白点.取3次制备的样品间的共同差异点进行胶内酶解,经基质辅助激光电离解析飞行时间质谱(MALDI-TOF-TOF/MS)获得肽质量指纹图谱,用Mascot软件在Swiss-Prot和NCBI数据库中搜索匹配蛋白.结果:表达量有明显下降的11个点,经鉴定对应10个蛋白:4个属于H pylori的氧化还原系统,即过氧化氢酶、超氧化物双歧酶、硫氧环蛋白和环氧蛋白还原酶;5个蛋白与代谢有关,即脯氨酸肽酶(proline peptidase)、果糖磷酸氢盐醛缩酶(fructose-bisphosphate aldolase)、无机焦磷酸酶(inorganic pyrophosphatase)、3-含氧酸辅酶A转移酶亚单位B(3-oxoacid CoA-transferase subunit B)和延伸因子(elongation factor P);另1个蛋白为假想蛋白HPAG0942.结论:H pylori在小鼠体内的定植适应过程中,伴有代谢酶表达的降低和抗氧化水平的减弱,提示代谢和抗氧化能力适度下调可能与菌株的定植力增强相关.

AIM： To analyze the protein profiles of mouseadapted H pylori SS1 and its initial strain 10700 and to find out the proteins associated with enhanced adhesive ability of SS1. METHODS： Three sets of cellular proteins of SS1 and 10 700 were prepared independently. The samples were separated by two-dimensional gel electrophoresis （2DE） technique, and the gels were scanned and recorded as digitalized images with high-resolution scanner. The images were analyzed by Image Master 2D software. Spots on gels were paired between 10 700 and SS1. Differently expressed proteins in all three sets were cut for in-gel digestion and identified by MALDI-TOF-TOF/MS. Peptide mass fingerprints were searched in the NCBI and Swiss-Prot database. RESULTS： Eleven down-regulated spots presented with ten proteins. Four were related to anti-oxidation, namely, catalase, thioredoxin reductase, superoxide dismutase and thioredoxin. Five were enzymes associated with metabolism, including proline peptidase, fructose- bisphosphate aldolase, inorganic pyrophosphatase, 3-oxoacid CoA-transferase subunit B and Elongation factor P. Another one was a putative protein HPAG0942. CONCLUSION： In the course of mouse-adapted, H pylori strain SS1 may increase its adhesive ability by decreasing metabolism and anti-oxidative level measurably.