ACS Publications. Most Trusted. Most Cited. Most Read
My Activity
CONTENT TYPES

Bfd, a New Class of [2Fe-2S] Protein That Functions in Bacterial Iron Homeostasis, Requires a Structural Anion Binding Site

  • Harshani Wijerathne
    Harshani Wijerathne
    Department of Chemistry, University of Kansas, Multidisciplinary Research Building, 2030 Becker Drive, Lawrence, Kansas 66047, United States
  • Huili Yao
    Huili Yao
    Department of Chemistry, Louisiana State University, 229A Choppin Hall, Baton Rouge, Louisiana 70803, United States
    More by Huili Yao
  • Yan Wang
    Yan Wang
    Department of Chemistry, University of Kansas, Multidisciplinary Research Building, 2030 Becker Drive, Lawrence, Kansas 66047, United States
    More by Yan Wang
  • Scott Lovell
    Scott Lovell
    Protein Structure Laboratory, Del Shankel Structural Biology Center, University of Kansas, 2034 Becker Drive, Lawrence, Kansas 66047, United States
    More by Scott Lovell
  • Kevin P. Battaile
    Kevin P. Battaile
    IMCA-CAT, Hauptman Woodward Medical Research Institute, 9700 South Cass Avenue, Building 435A, Argonne, Illinois 60439, United States
  • , and 
  • Mario Rivera*
    Mario Rivera
    Department of Chemistry, Louisiana State University, 229A Choppin Hall, Baton Rouge, Louisiana 70803, United States
    *Department of Chemistry, Louisiana State University, 229A Choppin Hall, Baton Rouge, LA 70803. Telephone: 225-578-1527. Fax: 225-578-3458. E-mail: [email protected]
    More by Mario Rivera
Cite this: Biochemistry 2018, 57, 38, 5533–5543
Publication Date (Web):September 5, 2018
https://doi.org/10.1021/acs.biochem.8b00823
Copyright © 2018 American Chemical Society

    Article Views

    913

    Altmetric

    -

    Citations

    LEARN ABOUT THESE METRICS
    Other access options
    Supporting Info (1)»

    Abstract

    Abstract Image

    Mobilization of iron from bacterioferritin (BfrB) requires specific interactions with a [2Fe-2S] ferredoxin (Bfd). Blocking the BfrB:Bfd interaction results in irreversible iron accumulation in BfrB and iron deficiency in the cytosol [Eshelman, K., et al. (2017) Metallomics9, 646–659]. The only known Bfd structure, which was obtained in complex with BfrB (Protein Data Bank entry 4E6K), indicated a new fold and suggested that the stability of Bfd is aided by an anion binding site consisting of R26, R29, and K46. We investigated the Bfd fold using site-directed mutagenesis, X-ray crystallography, and biochemistry in solution. The X-ray structure, which is nearly identical to that of Bfd in the BfrB:Bfd complex, shows that the [2Fe-2S] cluster preorganizes residues at the BfrB:Bfd interface into a structure complementary to the Bfd binding site on BfrB. Studies in solution showed rapid loss of the [2Fe-2S] cluster at a low ionic strength but higher stability with an increasing ionic strength, thus supporting a structural anion binding site. Structures of the R26E and R26E/K46Y mutants are nearly identical to that of Bfd, except for a new network of hydrogen bonds stabilizing the region encompassing the former anion binding site. The stability of the R26E and R26E/K46Y mutants, which is weakly and completely independent of solution ionic strength, respectively, corroborates that Bfd requires an anion binding site. The mutations, which caused only small changes to the strength of the BfrB:Bfd interaction and mobilization of iron from BfrB, indicate that the anion binding site in Bfd serves primarily a structural role.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Supporting Information

    ARTICLE SECTIONS
    Jump To

    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.biochem.8b00823.

    • Sequence alignment of Bfd, electronic absorption spectra of truncated and mutant Bfd, representative spectra obtained while monitoring the stability of truncated Bfd in Tris/NaCl buffers, and SPR data for the association between Bfd mutants and BfrB (PDF)

    Accession Codes

    Coordinates and structure factors were deposited as Protein Data Bank entries 6E6Q (truncated Bfd), 6E6R (truncated R26E Bfd), and 6E6S (truncated R26E/K46Y Bfd).

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

    Cited By

    This article is cited by 6 publications.

    1. Anabel Soldano, Huili Yao, Achala N. D. Punchi Hewage, Kevin Meraz, Joel K. Annor-Gyamfi, Richard A. Bunce, Kevin P. Battaile, Scott Lovell, Mario Rivera. Small Molecule Inhibitors of the Bacterioferritin (BfrB)–Ferredoxin (Bfd) Complex Kill Biofilm-Embedded Pseudomonas aeruginosa Cells. ACS Infectious Diseases 2021, 7 (1) , 123-140. https://doi.org/10.1021/acsinfecdis.0c00669
    2. Anabel Soldano, Huili Yao, Josephine R. Chandler, Mario Rivera. Inhibiting Iron Mobilization from Bacterioferritin in Pseudomonas aeruginosa Impairs Biofilm Formation Irrespective of Environmental Iron Availability. ACS Infectious Diseases 2020, 6 (3) , 447-458. https://doi.org/10.1021/acsinfecdis.9b00398
    3. Achala N. D. Punchi Hewage, Huili Yao, Baskar Nammalwar, Krishna Kumar Gnanasekaran, Scott Lovell, Richard A. Bunce, Kate Eshelman, Sahishna M. Phaniraj, Molly M. Lee, Blake R. Peterson, Kevin P. Battaile, Allen B. Reitz, Mario Rivera. Small Molecule Inhibitors of the BfrB–Bfd Interaction Decrease Pseudomonas aeruginosa Fitness and Potentiate Fluoroquinolone Activity. Journal of the American Chemical Society 2019, 141 (20) , 8171-8184. https://doi.org/10.1021/jacs.9b00394
    4. Mario Rivera. Mobilization of iron stored in bacterioferritin, a new target for perturbing iron homeostasis and developing antibacterial and antibiofilm molecules. Journal of Inorganic Biochemistry 2023, 247 , 112306. https://doi.org/10.1016/j.jinorgbio.2023.112306
    5. Minliang Guo, Miaomiao Gao, Jinjing Liu, Nan Xu, Hao Wang. Bacterioferritin nanocage: Structure, biological function, catalytic mechanism, self-assembly and potential applications. Biotechnology Advances 2022, 61 , 108057. https://doi.org/10.1016/j.biotechadv.2022.108057
    6. Huili Yao, Anabel Soldano, Leo Fontenot, Fabrizio Donnarumma, Scott Lovell, Josephine R. Chandler, Mario Rivera. Pseudomonas aeruginosa Bacterioferritin Is Assembled from FtnA and BfrB Subunits with the Relative Proportions Dependent on the Environmental Oxygen Availability. Biomolecules 2022, 12 (3) , 366. https://doi.org/10.3390/biom12030366

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    MENDELEY PAIRING EXPIRED
    Your Mendeley pairing has expired. Please reconnect