Structural fold, conservation and Fe(II) binding of the intracellular domain of prokaryote FeoB

Kuo Wei Hung, Yi Wei Chang, Edward T. Eng, Jai Hui Chen, Yi Chung Chen, Yuh Ju Sun, Chwan Deng Hsiao, Gang Dong, Krasimir A. Spasov, Vinzenz M. Unger, Tai huang Huang

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

FeoB is a G-protein coupled membrane protein essential for Fe(II) uptake in prokaryotes. Here, we report the crystal structures of the intracellular domain of FeoB (NFeoB) from Klebsiella pneumoniae (KpNFeoB) and Pyrococcus furiosus (PfNFeoB) with and without bound ligands. In the structures, a canonical G-protein domain (G domain) is followed by a helical bundle domain (S-domain), which despite its lack of sequence similarity between species is structurally conserved. In the nucleotide-free state, the G-domain's two switch regions point away from the binding site. This gives rise to an open binding pocket whose shallowness is likely to be responsible for the low nucleotide-binding affinity. Nucleotide binding induced significant conformational changes in the G5 motif which in the case of GMPPNP binding was accompanied by destabilization of the switch I region. In addition to the structural data, we demonstrate that Fe(II)-induced foot printing cleaves the protein close to a putative Fe(II)-binding site at the tip of switch I, and we identify functionally important regions within the S-domain. Moreover, we show that NFeoB exists as a monomer in solution, and that its two constituent domains can undergo large conformational changes. The data show that the S-domain plays important roles in FeoB function.

Original languageEnglish
Pages (from-to)501-512
Number of pages12
JournalJournal of Structural Biology
Volume170
Issue number3
DOIs
Publication statusPublished - 2010 Jun 1

Fingerprint

Nucleotides
GTP-Binding Proteins
Binding Sites
Pyrococcus furiosus
Printing
Klebsiella pneumoniae
Foot
Membrane Proteins
Ligands
Proteins
Protein Domains

Keywords

  • Feo
  • FeoB
  • Ferrous iron transport
  • G-protein
  • Klebsiella pneumoniae
  • Pyrococcus furiosus

ASJC Scopus subject areas

  • Structural Biology

Cite this

Structural fold, conservation and Fe(II) binding of the intracellular domain of prokaryote FeoB. / Hung, Kuo Wei; Chang, Yi Wei; Eng, Edward T.; Chen, Jai Hui; Chen, Yi Chung; Sun, Yuh Ju; Hsiao, Chwan Deng; Dong, Gang; Spasov, Krasimir A.; Unger, Vinzenz M.; Huang, Tai huang.

In: Journal of Structural Biology, Vol. 170, No. 3, 01.06.2010, p. 501-512.

Research output: Contribution to journalArticle

Hung, KW, Chang, YW, Eng, ET, Chen, JH, Chen, YC, Sun, YJ, Hsiao, CD, Dong, G, Spasov, KA, Unger, VM & Huang, TH 2010, 'Structural fold, conservation and Fe(II) binding of the intracellular domain of prokaryote FeoB', Journal of Structural Biology, vol. 170, no. 3, pp. 501-512. https://doi.org/10.1016/j.jsb.2010.01.017
Hung, Kuo Wei ; Chang, Yi Wei ; Eng, Edward T. ; Chen, Jai Hui ; Chen, Yi Chung ; Sun, Yuh Ju ; Hsiao, Chwan Deng ; Dong, Gang ; Spasov, Krasimir A. ; Unger, Vinzenz M. ; Huang, Tai huang. / Structural fold, conservation and Fe(II) binding of the intracellular domain of prokaryote FeoB. In: Journal of Structural Biology. 2010 ; Vol. 170, No. 3. pp. 501-512.
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AU - Hsiao, Chwan Deng

AU - Dong, Gang

AU - Spasov, Krasimir A.

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AB - FeoB is a G-protein coupled membrane protein essential for Fe(II) uptake in prokaryotes. Here, we report the crystal structures of the intracellular domain of FeoB (NFeoB) from Klebsiella pneumoniae (KpNFeoB) and Pyrococcus furiosus (PfNFeoB) with and without bound ligands. In the structures, a canonical G-protein domain (G domain) is followed by a helical bundle domain (S-domain), which despite its lack of sequence similarity between species is structurally conserved. In the nucleotide-free state, the G-domain's two switch regions point away from the binding site. This gives rise to an open binding pocket whose shallowness is likely to be responsible for the low nucleotide-binding affinity. Nucleotide binding induced significant conformational changes in the G5 motif which in the case of GMPPNP binding was accompanied by destabilization of the switch I region. In addition to the structural data, we demonstrate that Fe(II)-induced foot printing cleaves the protein close to a putative Fe(II)-binding site at the tip of switch I, and we identify functionally important regions within the S-domain. Moreover, we show that NFeoB exists as a monomer in solution, and that its two constituent domains can undergo large conformational changes. The data show that the S-domain plays important roles in FeoB function.

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