Angerer A, Klupp B, Braun V
Mikrobiologie II, Universität Tübingen, Germany.
J Bacteriol. 1992 Feb;174(4):1378-87. doi: 10.1128/jb.174.4.1378-1387.1992.
Serratia marcescens W225 expresses an unconventional iron(III) transport system. Uptake of Fe3+ occurs in the absence of an iron(III)-solubilizing siderophore, of an outer membrane receptor protein, and of the TonB and ExbBD proteins involved in outer membrane transport. The three SfuABC proteins found to catalyze iron(III) transport exhibit the typical features of periplasmic binding-protein-dependent systems for transport across the cytoplasmic membrane. In support of these conclusions, the periplasmic SfuA protein bound iron chloride and iron citrate but not ferrichrome, as shown by protection experiments against degradation by added V8 protease. The cloned sfuABC genes conferred upon an Escherichia coli aroB mutant unable to synthesize its own enterochelin siderophore the ability to grow under iron-limiting conditions (in the presence of 0.2 mM 2.2'-dipyridyl). Under extreme iron deficiency (0.4 mM 2.2'-dipyridyl), however, the entry rate of iron across the outer membrane was no longer sufficient for growth. Citrate had to be added in order for iron(III) to be translocated as an iron citrate complex in a FecA- and TonB-dependent manner through the outer membrane and via SfuABC across the cytoplasmic membrane. FecA- and TonB-dependent iron transport across the outer membrane could be clearly correlated with a very low concentration of iron in the medium. Expression of the sfuABC genes in E. coli was controlled by the Fur iron repressor gene. S. marcescens W225 was able to synthesize enterochelin and take up iron(III) enterochelin. It contained an iron(III) aerobactin transport system but lacked aerobactin synthesis. This strain was able to utilize the hydroxamate siderophores ferrichrome, coprogen, ferrioxamine B, rhodotorulic acid, and schizokinen as sole iron sources and grew on iron citrate as well. In contrast to E. coli K-12, S. marcescens could utilize heme. DNA fragments of the E. coli fhuA, iut, exbB, and fur genes hybridized with chromosomal S. marcescens DNA fragments, whereas no hybridization was obtained between S. marcescens chromosomal DNA and E. coli fecA, fhuE, and tonB gene fragments. The presence of multiple iron transport systems was also indicated by the increased synthesis of at least five outer membrane proteins (in the molecular weight range of 72,000 to 87,000) after growth in low-iron media. Serratia liquefaciens and Serratia ficaria produced aerobactin, showing that this siderophore also occurs in the genus Serratia.
粘质沙雷氏菌W225表达一种非常规的铁(III)转运系统。在没有铁(III)溶解铁载体、外膜受体蛋白以及参与外膜转运的托蛋白B(TonB)和ExbBD蛋白的情况下,Fe3+仍能被摄取。发现催化铁(III)转运的三种SfuABC蛋白表现出依赖周质结合蛋白的系统在跨细胞质膜转运方面的典型特征。支持这些结论的是,通过添加V8蛋白酶进行的抗降解保护实验表明,周质SfuA蛋白能结合氯化铁和柠檬酸铁,但不能结合铁色素。克隆的sfuABC基因赋予了一株无法合成自身肠螯合铁载体的大肠杆菌aroB突变体在铁限制条件下(在存在0.2 mM 2,2'-联吡啶的情况下)生长的能力。然而,在极端缺铁(0.4 mM 2,2'-联吡啶)的情况下,铁跨外膜的进入速率不再足以支持生长。必须添加柠檬酸盐,以便铁(III)以柠檬酸铁复合物的形式通过外膜以FecA和托蛋白B依赖的方式转运,并通过SfuABC跨细胞质膜转运。FecA和托蛋白B依赖的铁跨外膜转运与培养基中极低的铁浓度明显相关。sfuABC基因在大肠杆菌中的表达受铁调节蛋白Fur基因控制。粘质沙雷氏菌W225能够合成肠螯合铁并摄取铁(III)肠螯合铁。它含有一个铁(III)气杆菌素转运系统,但缺乏气杆菌素合成。该菌株能够利用异羟肟酸铁载体铁色素、粪生绿素、去铁胺B、玫红酸和裂殖菌素作为唯一铁源,也能在柠檬酸铁上生长。与大肠杆菌K-12不同,粘质沙雷氏菌能够利用血红素。大肠杆菌fhuA、iut、exbB和fur基因的DNA片段与粘质沙雷氏菌染色体DNA片段杂交,而粘质沙雷氏菌染色体DNA与大肠杆菌fecA、fhuE和tonB基因片段之间未获得杂交。在低铁培养基中生长后,至少五种外膜蛋白(分子量在72,000至87,000范围内)合成增加,这也表明存在多种铁转运系统。液化沙雷氏菌和无花果沙雷氏菌产生气杆菌素,表明这种铁载体也存在于沙雷氏菌属中。