title: FeoC from Klebsiella pneumoniae contains a [4Fe-4S] cluster.
abstract: Iron is essential for pathogen survival, virulence, and colonization. Feo is suggested to function as the ferrous iron (Fe(2+)) transporter. The enterobacterial Feo system is composed of 3 proteins: FeoB is the indispensable component and is a large membrane protein likely to function as a permease; FeoA is a small Src homology 3 (SH3) domain protein that interacts with FeoB; FeoC is a winged-helix protein containing 4 conserved Cys residues in a sequence suitable for harboring a putative iron-sulfur (Fe-S) cluster. The presence of an iron-sulfur cluster on FeoC has never been shown experimentally. We report that under anaerobic conditions, the recombinant Klebsiella pneumoniae FeoC (KpFeoC) exhibited hyperfine-shifted nuclear magnetic resonance (NMR) and a UV-visible (UV-Vis) absorbance spectrum characteristic of a paramagnetic center. The electron paramagnetic resonance (EPR) and extended X-ray absorption fine structure (EXAFS) results were consistent only with the [4Fe-4S] clusters. Substituting the cysteinyl sulfur with oxygen resulted in significantly reduced cluster stability, establishing the roles of these cysteines as the ligands for the Fe-S cluster. When exposed to oxygen, the [4Fe-4S] cluster degraded to [3Fe-4S] and eventually disappeared. We propose that KpFeoC may regulate the function of the Feo transporter through the oxygen- or iron-sensitive coordination of the Fe-S cluster.
authors: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
authors: Kuang-Lung
authors: KL
authors: Hsueh
authors: Liang-Kun
authors: LK
authors: Yu
authors: Yung-Han
authors: YH
authors: Chen
authors: Ya-Hsin
authors: YH
authors: Cheng
authors: Yin-Cheng
authors: YC
authors: Hsieh
authors: Shyue-chu
authors: SC
authors: Ke
authors: Kuo-Wei
authors: KW
authors: Hung
authors: Chun-Jung
authors: CJ
authors: Chen
authors: Tai-huang
authors: TH
authors: Huang
mesh_terms: D015502:Absorptiometry, Photon; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D007506:Iron-Sulfur Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D010084:Oxidation-Reduction
chemical_list: D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins
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