Zheng Jianhua, Wei Candong, Leng Wenchuan, Dong Jie, Li Rui, Li Weijun, Wang Jin, Zhang Zongde, Jin Qi
College of Biological Sciences, China Agricultural University, Beijing, PR China.
Proteomics. 2007 Nov;7(21):3919-31. doi: 10.1002/pmic.200700342.
Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine has been known for a long time to prevent tuberculosis (TB) worldwide since 1921. Nonetheless, we know little about BCG membrane proteome. In the present study, we utilized alkaline incubation and Triton X-114-based methods to enrich BCG membrane proteins and subsequently digested them using proteolytic enzyme. The recovered peptides were further separated by 2-D LC and identified by ESI-MS/MS. As a result, total 474 proteins were identified, including 78 integral membrane proteins (IMPs). Notably, 18 BCG IMPs were described for the first time in mycobacterium. Further analysis of the 78 IMPs indicated that the theoretical molecular mass distribution of them ranged from 8.06 to 167.86 kDa and pI scores ranged from 4.40 to 11.60. Functional classification revealed that a large proportion of the identified IMPs (67.9%, 53 out of 78) were involved in cell wall and cell processes functional group. In conclusion, here we reported a comprehensive profile of the BCG membrane subproteome. The present investigation may allow the identification of some valuable vaccine and drug target candidates and thus provide basement for future designing of preventive, diagnostic, and therapeutic strategies against TB.
自1921年以来,卡介苗(BCG)在全球范围内一直被用于预防结核病(TB),这已为人所知良久。尽管如此,我们对卡介苗的膜蛋白质组了解甚少。在本研究中,我们采用碱性孵育和基于Triton X-114的方法富集卡介苗膜蛋白,随后用蛋白酶对其进行消化。回收的肽段通过二维液相色谱进一步分离,并用电喷雾串联质谱(ESI-MS/MS)进行鉴定。结果,共鉴定出474种蛋白质,其中包括78种整合膜蛋白(IMP)。值得注意的是,有18种卡介苗IMP是在分枝杆菌中首次被描述的。对这78种IMP的进一步分析表明,它们的理论分子量分布范围为8.06至167.86 kDa,等电点分数范围为4.40至11.60。功能分类显示,所鉴定的IMP中有很大一部分(67.9%,78种中的53种)参与细胞壁和细胞过程功能组。总之,我们在此报告了卡介苗膜亚蛋白质组的全面概况。本研究可能有助于鉴定一些有价值的疫苗和药物候选靶点,从而为未来设计抗结核的预防、诊断和治疗策略提供基础。