血清2型猪链球菌全菌体蛋白的免疫蛋白质组学研究  

Immunoproteomic analysis of the whole cell lysate proteins in Streptococcus suis type 2

作　　者：罗霞[1] 孙强正[1] 肖迪[1] 叶长芸[1] 郑翰[1] 景怀琦[1] 徐建国[1] 

机构地区：[1]传染病预防控制重点实验室,中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《中国人兽共患病学报》2009年第4期299-303,共5页Chinese Journal of Zoonoses

基　　金：国际合作项目(No.2008DFA31830)资助

摘　　要：目的寻找血清2型猪链球菌(Streptococcus suis,S.suis)全菌体蛋白中具有抗原活性的蛋白,为S.suis特异诊断抗原和新的疫苗侯选抗原的筛选提供线索。方法采用免疫蛋白质组学技术,分析和研究血清2型S.suisSC84菌株全菌体蛋白中能与猪链球菌病人恢复期血清发生免疫杂交的蛋白点。应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)对这些蛋白进行鉴定。结果S.suisSC84全菌体蛋白与3份猪链球菌病人恢复期血清进行免疫杂交反应,发现8个与IgG抗体发生阳性反应的蛋白点。经MALDI-TOF-MS鉴定,这8个蛋白分别为溶菌酶释放蛋白(MRP)、琥珀酸脱氢酶/延胡索酸还原酶黄素蛋白亚单位(SDHFp/FRDFp)、触发因子(TF)、延长因子G(EF-G)、细菌糖磷酸转移酶系统甘露糖特异ⅡAB亚单位(PTS/MAN-IIAB)、丙酮酸激酶(PK)、6-磷酸果糖激酶(6-PFK)、色氨酸-tRNA合成酶(TrpRS)。MRP定位于胞壁;TF、PTS/MAN-IIAB、PK及TrpRS定位于胞质;SDHFp/FRDFp、EF-G及6-PFK定位于胞外。并且确定了它们在S.suis05ZYH33菌株基因组的编码基因。结论在S.suisSC84全菌体蛋白中发现8个抗原活性蛋白,其中SDHFp/FRDFp、TF、EF-G、PTS、PK、6-PFK及TrpRS是在S.suis新发现的抗原性蛋白,它们作为特异诊断抗原和疫苗侯选抗原的前景有待进一步研究。To search for the activated proteins with antigenic activity in the whole cell lysate of Streptococcus suis type 2 in order to provide some information to find out the proper candidates of the specific antigen for diagnosis and preparation of the new vaccine. An immunoproteomic assay was used to analyze the antigenic proteins from the whole cell lysate of S. suis type 2 SC84 strain which could be recognized by the convalescent sera of patients infected with this organisms as spots on the PVDF membrane. These proteins on duplicate gel were excised and identified by matrix-assisted laser desorption-ionization-time of flight mass spectrometry （MALDI-TOP-MS）. It was found that the whole cell lysate proteins of S. suis SC84 strain could react specifically with 3 samples of the convalescent sera of patients and 8 protein spots showing positive reactions with the IgG antibodies were obtained. They were named respectively as muramidase-released protein （MRP, locating at cell wall）; succinate dehydrogenase/fumarate reductase flavoprotein subunit （SDHFp/FRDFP, extraceliular）; bacterial trigger factor （TF, cytoplasmic） ; elongation factor G （EF-G, extracellular） ; PTS system sorbose subfamily Ⅰ（PTS/MAN-ⅡAB, cytoplasmic） ; pyruvate kinase （PK, cytoplasmic）; phosphofructokinase, （PK. extracellullar） and tRNA synthase class Ⅰ （cytoplasmic）. In addition, the coding genes for each identified protein could be demonstrated in the genome of S. suis SS2 strain isolated from an outbreak of infection in Sichuan province of China in 2005. Most of these proteins can be developed as the suitable candidates for the bacterial diagnosis as well as for the preparation of new vaccine.

关 键 词：血清2型猪链球菌 全菌体蛋白 免疫蛋白质组学 

分 类 号：R378[医药卫生—病原生物学]