Meyer J M, Neely A, Stintzi A, Georges C, Holder I A
Laboratoire de Microbiologie et de Génétique, Université Louis-Pasteur, Strasbourg, France.
Infect Immun. 1996 Feb;64(2):518-23. doi: 10.1128/iai.64.2.518-523.1996.
The role of pyoverdin, the main siderophore in iron-gathering capacity produced by Pseudomonas aeruginosa, in bacterial growth in vivo is controversial, although iron is important for virulence. To determine the ability of pyoverdin to compete for iron with the human iron-binding protein transferrin, wild-type P. aeruginosa ATCC 15692 (PAO1 strain) and PAO pyoverdin-deficient mutants were grown at 37 degrees C in bicarbonate-containing succinate medium to which apotransferrin had been added. Growth of the pyoverdin-deficient mutants was fully inhibited compared with that of the wild type but was restored when pyoverdin was added to the medium. Moreover, when growth took place at a temperature at which no pyoverdin production occurred (43 degrees C), the wild-type PAO1 strain behaved the same as the pyoverdin-deficient mutants, with growth inhibited by apotransferrin in the presence of bicarbonate and restored by pyoverdin supplementation. Growth inhibition was never observed in bicarbonate-free succinate medium, whatever the strain and the temperature for growth. In vivo, in contrast to results obtained with the wild-type strain, pyoverdin-deficient mutants demonstrated no virulence when injected at 10(2) CFU into burned mice. However, virulence was restored when purified pyoverdin originating from the wild-type strain was supplemented during the infection. These results strongly suggest that pyoverdin competes directly with transferrin for iron and that it is an essential element for in vivo iron gathering and virulence expression in P. aeruginosa. Rapid removal of iron from [59Fe]ferritransferrin by pyoverdin in vitro supports this view.
尽管铁对铜绿假单胞菌的毒力很重要,但该菌产生的主要铁载体——绿脓菌素在体内细菌生长中的作用仍存在争议。为了确定绿脓菌素与人类铁结合蛋白转铁蛋白竞争铁的能力,将野生型铜绿假单胞菌ATCC 15692(PAO1菌株)和PAO绿脓菌素缺陷型突变体在含有碳酸氢盐的琥珀酸盐培养基中于37℃培养,该培养基中已添加了脱铁转铁蛋白。与野生型相比,绿脓菌素缺陷型突变体的生长完全受到抑制,但当向培养基中添加绿脓菌素时,生长得以恢复。此外,当在不产生绿脓菌素的温度(43℃)下生长时,野生型PAO1菌株的表现与绿脓菌素缺陷型突变体相同,在有碳酸氢盐存在的情况下,脱铁转铁蛋白会抑制其生长,而补充绿脓菌素后生长恢复。无论菌株和生长温度如何,在无碳酸氢盐的琥珀酸盐培养基中从未观察到生长抑制现象。在体内,与野生型菌株的结果相反,将绿脓菌素缺陷型突变体以10²CFU注射到烧伤小鼠体内时,未表现出毒力。然而,在感染期间补充源自野生型菌株的纯化绿脓菌素后,毒力得以恢复。这些结果强烈表明,绿脓菌素直接与转铁蛋白竞争铁,并且它是铜绿假单胞菌体内铁摄取和毒力表达的必需元素。绿脓菌素在体外能快速从[⁵⁹Fe]铁转铁蛋白中去除铁,这支持了这一观点。