Braud Armelle, Hoegy Françoise, Jezequel Karine, Lebeau Thierry, Schalk Isabelle J
Métaux et Microorganismes, Chimie, Biologie et Applications, UMR 7175-LC1, CNRS-Université Louis Pasteur, ESBS, Illkirch, Strasbourg, France.
Environ Microbiol. 2009 May;11(5):1079-91. doi: 10.1111/j.1462-2920.2008.01838.x. Epub 2009 Jan 7.
Pyoverdine (PvdI) is the major siderophore secreted by Pseudomonas aeruginosa PAOI in order to get access to iron. After being loaded with iron in the extracellular medium, PvdI is transported across the bacterial outer membrane by the transporter, FpvAI. We used the spectral properties of PvdI to show that in addition to Fe(3+), this siderophore also chelates, but with lower efficiencies, all the 16 metals used in our screening. Afterwards, FpvAI at the cell surface binds Ag(+), Al(3+), Cd(2+), Co(2+), Cu(2+), Fe(3+), Ga(3+), Hg(2+), Mn(2+), Ni(2+) or Zn(2+) in complex with PvdI. We used Inductively Coupled Plasma-Atomic Emission Spectrometry to monitor metal uptake in P. aeruginosa: TonB-dependent uptake, in the presence of PvdI, was only efficient for Fe(3+). Cu(2+), Ga(3+), Mn(2+) and Ni(2+) were also transported into the cell but with lower uptake rates. The presence of Al(3+), Cu(2+), Ga(3+), Mn(2+), Ni(2+) and Zn(2+) in the extracellular medium induced PvdI production in P. aeruginosa. All these data allow a better understanding of the behaviour of the PvdI uptake pathway in the presence of metals other than iron: FpvAI at the cell surface has broad metal specificity at the binding stage and it is highly selective for Fe(3+) only during the uptake process.
绿脓菌素(PvdI)是铜绿假单胞菌PAO1分泌的主要铁载体,用于获取铁元素。在细胞外培养基中装载铁后,PvdI通过转运蛋白FpvAI穿过细菌外膜。我们利用PvdI的光谱特性表明,除了Fe(3+)外,这种铁载体还能螯合我们筛选中使用的所有16种金属,但效率较低。之后,细胞表面的FpvAI与与PvdI络合的Ag(+)、Al(3+)、Cd(2+)、Co(2+)、Cu(2+)、Fe(3+)、Ga(3+)、Hg(2+)、Mn(2+)、Ni(2+)或Zn(2+)结合。我们使用电感耦合等离子体原子发射光谱法监测铜绿假单胞菌中的金属摄取:在存在PvdI的情况下,TonB依赖性摄取仅对Fe(3+)有效。Cu(2+)、Ga(3+)、Mn(2+)和Ni(2+)也被转运到细胞中,但摄取率较低。细胞外培养基中Al(3+)、Cu(2+)、Ga(3+)、Mn(2+)、Ni(2+)和Zn(2+)的存在会诱导铜绿假单胞菌产生PvdI。所有这些数据有助于更好地理解在除铁以外的金属存在下PvdI摄取途径的行为:细胞表面的FpvAI在结合阶段具有广泛的金属特异性,而在摄取过程中仅对Fe(3+)具有高度选择性。