Guidi-Rontani C, Collier R J
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Mol Microbiol. 1987 Jul;1(1):67-72. doi: 10.1111/j.1365-2958.1987.tb00528.x.
We have constructed defined deletions in the structural gene of Pseudomonas aeruginosa exotoxin A (ETA) in order to probe the function of Domain I of this protein. Three forms of the gene containing specific deletions were expressed in a strain of Escherichia coli K12 with lesions in the htpR and Ion genes; extracts containing the gene products were tested for ADP-ribosylation activity, cytotoxicity, and ability to protect sensitive cells from the cytotoxic action of authentic ETA. Two of the mutant ETAs gave concentration-dependent protection against authentic ETA, and protection correlated with the presence of the bulk of Domain I. The results support the notion that Domain I functions in binding the toxin to specific cell-surface receptors.
我们构建了铜绿假单胞菌外毒素A(ETA)结构基因的特定缺失,以探究该蛋白结构域I的功能。三种含有特定缺失的基因形式在htpR和Ion基因有损伤的大肠杆菌K12菌株中表达;对含有基因产物的提取物进行ADP-核糖基化活性、细胞毒性以及保护敏感细胞免受天然ETA细胞毒性作用能力的测试。两种突变型ETA对天然ETA提供浓度依赖性保护,且保护作用与大部分结构域I的存在相关。结果支持结构域I在将毒素结合到特定细胞表面受体中发挥作用这一观点。