Identification of a novel iron regulated staphylococcal surface protein with haptoglobin‐haemoglobin binding activity
Open accessAbstract
Summary: Staphylococcus aureus is an extremely adaptable pathogen causing a wide variety of infections. Staphylococcal surface proteins that directly interact with host extracellular proteins greatly contribute to virulence and are involved in adhesion, immune escape and nutrient acquisition. In our extensive search for highly immunogenic, in vivo‐expressed, staphylococcal proteins, previously, we identified a novel member of the family of Gram‐positive anchor motif proteins with a predicted 895 amino acid long sequence. In order to determine the ligand for this novel LPXTG cell wall protein, we employed affinity purification of human plasma using the recombinant form of the protein. Two‐dimensional electrophoresis of eluted plasma proteins identified haptoglobin as a specific binding partner. Importantly, we also observed this specific ligand binding when living S. aureus cells were exposed to biotin‐labelled haptoglobin (Hp) in a FACS‐based assay. Targeted deletion of the gene eliminated Hp‐binding, a function that has not been attributed to S. aureus before. Based on these data we specified the protein as the staphylococcal ha ptoglobin r eceptor A (HarA). Similarly to other haptoglobin receptors identified in Gram‐negative pathogens, HarA binds not only Hp, but also haptoglobin‐haemoglobin complexes with an even higher affinity, as demonstrated in in vitro binding assays. Employing specific deletion mutants, ligand binding was localized to two homologous regions with about 145 amino acid residues located within the N‐terminal part of the protein. In addition, we demonstrated that expression of HarA was strictly controlled by iron through the iron‐dependent transcriptional regulator Fur. Based on these data we propose that HarA can be added to the list of staphylococcal virulence factors with a most likely function related to iron acquisition.
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IsdH contains three NEAT domains; NEAT domain 1 binds hemoglobin and haptoglobin, NEAT domain 2 binds hemoglobin with nanomolar affinity, and NEAT domain 3 weakly binds heme.
“IsdH contains three NEAT domains (25). Structural analysis of the IsdH NEAT domains revealed that NEAT domain 1 binds both hemoglobin and haptoglobin. NEAT domain 2 binds hemoglobin with nanomolar affinities and NEAT domain 3 weakly binds heme.”
From Molecular Mechanisms of <i>Staphylococcus aureus</i> Iron Acquisition (2011) · cites this paper as [25]
The haptoglobin-hemoglobin complex is a viable iron source for S. aureus and is bound by IsdH.
“The haptoglobin-hemoglobin complex is also a viable source of iron for S. aureus and this complex is bound by IsdH (25, 27, 78) (Fig 1. B).”
From Molecular Mechanisms of <i>Staphylococcus aureus</i> Iron Acquisition (2011) · cites this paper as [25]
IsdH contains three NEAT domains; N1 and N2 bind both Hb and Hp, and N3 binds heme.
“IsdH contains three NEAT domains, N1 and N2 bind both Hb and Hp, andN3bindsheme [109,110].”
From Heme Synthesis and Acquisition in Bacterial Pathogens (2016) · cites this paper as [109]
IsdH binds hemoglobin and heme in vitro.
“IsdH is another protein that binds hemoglobin and heme in vitro [18, 19].”
From IsdB-dependent hemoglobin binding is required for acquisition of heme by Staphylococcus aureus (2014) · cites this paper as [19]
Predicted physiological concentrations of free heme and cell-free plasma hemoglobin are 30 nM and 150 nM, respectively.
“consistent with the predicted physiological concentrations of free heme and cell-free plasma hemoglobin of 30 and 150 nM, respectively [19, 39].”
From IsdB-dependent hemoglobin binding is required for acquisition of heme by Staphylococcus aureus (2014) · cites this paper as [19]
The authors' data suggest that Isd is a high-affinity heme-iron acquisition system most useful at low nanomolar heme/hemoglobin concentrations.
“Our data suggest that Isd is a high-af fi nity heme-iron acquisition system that has maximal utility at the low nanomolar concentrations of heme/hemoglobin, consistent with the predicted physiological concentrations of free heme and cell-free plasma hemoglobin of 30 and 150 nM, respectively [19, 39].”
From IsdB-dependent hemoglobin binding is required for acquisition of heme by Staphylococcus aureus (2014) · cites this paper as [19]
S. aureus binds haptoglobin-hemoglobin complexes in vitro, suggesting it uses this complex as a nutrient iron source.
“S. aureus binds haptoglobin-hemoglobin complexes in vitro, suggesting that this protein complex is exploited as a source of nutrient iron [8].”
From A battle for iron: host sequestration and Staphylococcus aureus acquisition (2012) · cites this paper as [8]
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- Authors
- Dryla, A., Gelbmann, D., Von Gabain, A., & Nagy, E.
- Affiliations
- Intercell (Austria) (AT)
- Corresponding author
- Nagy, Eszter
- ORCID
- Nagy, Eszter
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 2003-05-28
- Volume
- 49
- Issue
- 1
- Pages
- 37-53
- ISSN
- 0950-382X (ISSN-L)
- 1365-2958
- OpenAlex
- W2110043149
- Cited by
- 153
- References
- 9
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 3 · 2023: 1 · 2022: 5 · 2021: 4 · 2020: 6 · 2019: 9 · 2018: 4 · 2017: 4 · 2016: 6 · 2015: 6 · 2014: 10 · 2013: 9 · 2012: 13
- FWCI
- 5.67
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Microbiology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antimicrobial Resistance in Staphylococcus
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Staphylococcus aureus, haras, iron acquisition, iron regulation, virulence factors, host-pathogen interaction, ligand binding
- MeSH terms
- Amino Acid Sequence, Bacterial Proteins, Cell Division, Cell Wall, Haptoglobins, Hemoglobins, Humans, Iron, Membrane Proteins, Molecular Sequence Data, Protein Binding, Recombinant Proteins, Staphylococcus aureus, Sequence Alignment
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