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

Journal of Structural Biology
Kuo-Wei HungTai-Huang Huang

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 chan...Continue Reading

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Citations

May 15, 2012·Journal of Biomolecular NMR·Kuo-Wei HungTai Huang Huang
Nov 22, 2011·Acta Crystallographica. Section D, Biological Crystallography·Miriam Rose AshMika Jormakka
Dec 6, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Miriam Rose AshMika Jormakka
Dec 20, 2015·FEMS Microbiology Reviews·Cheryl K Y LauHans J Vogel
Jun 13, 2015·Protein Expression and Purification·Gregor HageluekenOlav Schiemann
Jul 6, 2014·Protein Expression and Purification·Saeed SeyedmohammadHenrietta Venter
Jun 29, 2018·Metallomics : Integrated Biometal Science·Alexandrea E SestokAaron T Smith
Aug 21, 2013·Journal of Bacteriology·Kuang-Lung HsuehTai-huang Huang
Feb 3, 2016·Journal of Bacteriology·Begoña StevensonShelley M Payne
Dec 19, 2019·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Gabriel Soares Guerra, Andrea Balan
Apr 3, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Chandrika N DeshpandeMika Jormakka

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