双向电泳和肽质谱技术分析结核杆菌H37Ra感染巨噬细胞引致的差异表达蛋白

Analysis of the Different Expression Proteins of Macrophages Infected with Mycobacterium Tuberculosis H37Ra by 2-D Electrophoresis and Mass Spectrum

利用双向电泳及质谱技术对感染结核杆菌H37Ra株前后的巨噬细胞U937株的蛋白质组进行分离和比较分析.双向电泳后在分子质量20～200ku,等电点pI3～10范围内分离出1913个不同蛋白质斑点,肉眼可以明显看见4个蛋白质斑点在表达上的差异.对其中分子质量约75ku等电点约9.5的蛋白质斑点酶解后,进行质谱分析,鉴定为分子质量75.4ku、等电点9.52的蛋白DEAD/HBoxPolypeptide18.说明用蛋白质组学研究结核杆菌与巨噬细胞相互作用机理是可行的,所鉴定的蛋白质DEAD/HBoxPolypeptide18可能与巨噬细胞防御系统有关.

Analysis of the different expression proteins of macrophages infected with mycobacterium tuberculosis H37Ra by 2D electrophoresis and mass spectrum. In the rang of molecular mass from 20 to 200 ku, pI 310, there are about 1 900 different protein spots by computer analysis. By the naked eyes there are 4 protein spots with quantity difference. One protein spot with molecular mass about 75 ku pI 9.5 was selected to be cut and identified by MALDITOFMS. This protein is DEAD/H Box Polypeptide 18 with molecular mass 75.4 ku, pI 9.52. It is feasible for use in proteomics to study the different proteins on the interaction of macrophages and mycobacterium tuberculosis. The protein DEAD/H Box Polypeptide 18 identified may play a role in the defence system of macrophages against mycobacterium tuberculosis.