Novel Mechanism of Hemin Capture by Hbp2, the Hemoglobin-binding Hemophore from Listeria monocytogenes
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Iron is an essential nutrient that is required for the growth of the bacterial pathogen Listeria monocytogenes. In cell cultures, this microbe secretes hemin/hemoglobin-binding protein 2 (Hbp2; Lmo2185) protein, which has been proposed to function as a hemophore that scavenges heme from the environment. Based on its primary sequence, Hbp2 contains three NEAr transporter (NEAT) domains of unknown function. Here we show that each of these domains mediates high affinity binding to ferric heme (hemin) and that its N- and C-terminal domains interact with hemoglobin (Hb). The results of hemin transfer experiments are consistent with Hbp2 functioning as an Hb-binding hemophore that delivers hemin to other Hbp2 proteins that are attached to the cell wall. Surprisingly, our work reveals that the central NEAT domain in Hbp2 binds hemin even though its primary sequence lacks a highly conserved YXXXY motif that is used by all other previously characterized NEAT domains to coordinate iron in the hemin molecule. To elucidate the mechanism of hemin binding by Hbp2, we determined crystal structures of its central NEAT domain (Hbp2(N2); residues 183-303) in its free and hemin-bound states. The structures reveal an unprecedented mechanism of hemin binding in which Hbp2(N2) undergoes a major conformational rearrangement that facilitates metal coordination by a non-canonical tyrosine residue. These studies highlight previously unrecognized plasticity in the hemin binding mechanism of NEAT domains and provide insight into how L. monocytogenes captures heme iron.
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Many other pathogens have evolved NEAT-containing heme acquisition systems, including Staphylococcus lugdunensis, L. monocytogenes, and Streptococcus pyogenes.
“In addition to S. aureus and B. anthracis, many other pathogens have evolved NEAT-containing heme acquisition systems, including Staphylococcus lugdunensis, L. monocytogenes, and Streptococcus pyogenes [136–143].”
From Heme Synthesis and Acquisition in Bacterial Pathogens (2016) · cites this paper as [139]
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- Authors
- Malmirchegini, G. R., Sjodt, M., Shnitkind, S., Sawaya, M. R., Rosinski, J., Newton, S. M., Klebba, P. E., & Clubb, R. T.
- Affiliations
- University of California, Los Angeles (US)
- Howard Hughes Medical Institute (US)
- Kansas State University (US)
- Corresponding author
- Clubb, Robert T.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2014-12
- Volume
- 289
- Issue
- 50
- Pages
- 34886-34899
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC4263887
- OpenAlex
- W2000991144
- Cited by
- 33
- References
- 58
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 4 · 2023: 1 · 2022: 4 · 2021: 1 · 2019: 3 · 2018: 2 · 2017: 2 · 2016: 5 · 2015: 8
- FWCI
- 2.49
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- eScholarship (California Digital Library) (submittedVersion, cc-by)
- Funders
- National Institutes of Health (AI52217, GM53836, F31GM101931)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- hemophore, Listeria monocytogenes, hemin binding, NEAT domain, heme acquisition, iron acquisition, hemoglobin binding, conformational change, crystal structure, bacterial virulence factors
- MeSH terms
- Amino Acid Sequence, Bacterial Proteins, Hemin, Hemoglobins, Humans, Kinetics, Listeria monocytogenes, Models, Molecular, Molecular Sequence Data, Protein Binding, Protein Structure, Tertiary, Crystallography, X-Ray
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