Kuroda K, Kageyama M
J Biochem. 1981 Jun;89(6):1721-36. doi: 10.1093/oxfordjournals.jbchem.a133372.
Pseudomonas aeruginosa strain PAF41 was found to produce a new F-type pyocin, pyocin F3, the action spectrum of which was different from those of previously reported pyocins F1 and F2. These three F-type pyocins were compared with respect to their structure and biological properties. These pyocins were almost the same with regard to the structure and the dimensions, and have similar amino acid compositions and S values. The particle weights of these pyocins were also suggested to be similar. Analyses of subunit proteins by SDS-polyacrylamide slab gel electrophoresis showed that these pyocins were composed of 5 major (bands 1, 2, 3, 4, and 6) and 2 minor (bands 5 and 7) subunit proteins and that no difference in the mobilities of these subunit proteins could be detected among the pyocins except that of the second major subunit protein (band 4), which did differ. Pyocins F1, F2, and F3 were immunologically cross-reactive, and carried common antigens as well as specific ones. It was shown that band 6 was a common antigen among the three pyocins and that band 4 was antigenically different in pyocins F1 and F3 by immunological reaction after protein blotting. Electron microscopic observation of pyocin particles treated with anti-sera revealed that the common antigens were located on the rod part and the specific ones were on the fiber part. Pyocin F3 was neutralized by both anti-F3 and anti-F1 sera showing apparent first order rate kinetics, whereas the neutralization for pyocin F1 by these sera did not show such kinetics, but a considerable increment of pyocin F1 activity was observed when small amounts of the sera were added. The increment seemed to be due to the antibodies common to pyocins F1, F2, and F3. A phage, which had a flexuous rod-like tail, was found to be immunologically cross-reactive with the three pyocins and was named KF1.
发现铜绿假单胞菌PAF41菌株可产生一种新的F型绿脓菌素,即绿脓菌素F3,其作用谱与先前报道的绿脓菌素F1和F2不同。对这三种F型绿脓菌素的结构和生物学特性进行了比较。这些绿脓菌素在结构和尺寸方面几乎相同,具有相似的氨基酸组成和S值。还表明这些绿脓菌素的颗粒重量相似。通过SDS-聚丙烯酰胺平板凝胶电泳对亚基蛋白进行分析表明,这些绿脓菌素由5种主要亚基蛋白(条带1、2、3、4和6)和2种次要亚基蛋白(条带5和7)组成,除了第二种主要亚基蛋白(条带4)的迁移率存在差异外,在这些绿脓菌素之间未检测到这些亚基蛋白迁移率的差异。绿脓菌素F1、F2和F3具有免疫交叉反应性,携带共同抗原以及特异性抗原。蛋白质印迹后的免疫反应表明,条带6是三种绿脓菌素中的共同抗原,而条带4在绿脓菌素F1和F3中抗原性不同。用抗血清处理绿脓菌素颗粒后的电子显微镜观察表明,共同抗原位于杆状部分,特异性抗原位于纤维部分。绿脓菌素F3可被抗F3血清和抗F1血清中和,呈现明显的一级速率动力学,而这些血清对绿脓菌素F1的中和未呈现这种动力学,但当加入少量血清时,观察到绿脓菌素F1活性有相当大的增加。这种增加似乎是由于绿脓菌素F1、F2和F3共有的抗体所致。发现一种具有弯曲杆状尾部的噬菌体与这三种绿脓菌素具有免疫交叉反应性,被命名为KF1。