Abstract
During infections Stahpylococcus aureus preferentially uses heme as an iron source, which it captures from human hemoglobin using the Iron regulated surface determinant (Isd) system. On the cell surface two related staphylococcal surface receptors called IsdH and IsdB bind to hemoglobin and extract its heme. Both receptors contain multiple NEAr iron Transporter (NEAT) domains that either bind to hemoglobin, or to heme. All previous structural studies have investigated individual NEAT domains and have not explored how the domains might interact with one another to synergistically extract heme from hemoglobin. Here, we report the near complete 1H, 13C and 15N backbone resonance assignments of a bi-domain unit from IsdH that contains the N2 and N3 NEAT domains, which bind to hemoglobin and heme, respectively (IsdHN2N3, residues 326–660, 39 kDa). The assigned backbone resonances lay the foundation for future NMR studies that will explore the molecular basis of IsdH function.
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References
Andrade MA, Ciccarelli FD, Perez-Iratxeta C, Bork P (2002) NEAT: a domain duplicated in genes near the components of a putative Fe3+ siderophore transporter from Gram-positive pathogenic bacteria. Genome Biol 3:RESEARCH0047
Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293
Dryla A, Gelbmann D, von Gabain A, Nagy E (2003) Identification of a novel iron regulated staphylococcal surface protein with haptoglobin-haemoglobin binding activity. Mol Microbiol 49:37–53
Grigg JC, Vermeiren CL, Heinrichs DE, Murphy ME (2007) Haem recognition by a Staphylococcus aureus NEAT domain. Mol Microbiol 63:139–149
Keller R (2005) Computer-Aided Resonance Assignment, version 1.2.3. http://cara.nmr.ch
Kumar KK, Jacques DA, Pishchany G, Caradoc-Davies T, Spirig T, Malmirchegini GR, Langley DB, Dickson DF, Mackay JP, Clubb RT, Skaar EP, Guss JM, Gell DA (2011) The structural basis for haemoglobin capture by Staphylococcus aureus. J Biol Chem 286:38439–38447
Maresso AW, Schneewind O (2006) Iron acquisition and transport in Staphylococcus aureus. Biometals 19:193–203
Pilpa RM, Fadeev EA, Villareal VA, Wong ML, Phillips M, Clubb RT (2006) Solution structure of the NEAT (NEAr Transporter) domain from IsdH/HarA: the human hemoglobin receptor in Staphylococcus aureus. J Mol Biol 360:435–447
Pilpa RM, Robson SA, Villareal VA, Wong ML, Phillips M, Clubb RT (2009) Functionally distinct NEAT (NEAr Transporter) domains within the Staphylococcus aureus IsdH/HarA protein extract heme from methemoglobin. J Biol Chem 284:1166–1176
Sharp KH, Schneider S, Cockayne A, Paoli M (2007) Crystal structure of the heme-IsdC complex, the central conduit of the Isd iron/heme uptake system in Staphylococcus aureus. J Biol Chem 282:10625–10631
Shen Y, Delaglio F, Cornilescu G, Bax A (2009) TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts. J Biomol NMR 44:213–223
Villareal VA, Pilpa RM, Robson SA, Fadeev EA, Clubb RT (2008) The IsdC protein from Staphylococcus aureus uses a flexible binding pocket to capture heme. J Biol Chem 283:31591–31600
Villareal VA, Spirig T, Robson SA, Liu M, Lei B, Clubb RT (2011) Transient weak protein–protein complexes transfer heme across the cell wall of Staphylococcus aureus. J Am Chem Soc 133:14176–14179
Watanabe M, Tanaka Y, Suenaga A, Kuroda M, Yao M, Watanabe N, Arisaka F, Ohta T, Tanaka I, Tsumoto K (2008) Structural basis for multimeric heme complexation through a specific protein-heme interaction: the case of the third neat domain of IsdH from Staphylococcus aureus. J Biol Chem 283:28649–28659
Acknowledgments
We would like to thank Reza Malmirchegini for help with DNA cloning, Dr. Robert Peterson for assistance with NMR experiments and Dr. Fred Damberger for useful discussions. This work was supported by National Institutes of Health Grant AI52217 to Robert T. Clubb. Thomas Spirig was supported in part by Swiss National Science Foundation Fellowship PBEZP3-124281.
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Spirig, T., Clubb, R.T. Backbone 1H, 13C and 15N resonance assignments of the 39 kDa staphylococcal hemoglobin receptor IsdH. Biomol NMR Assign 6, 169–172 (2012). https://doi.org/10.1007/s12104-011-9348-8
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DOI: https://doi.org/10.1007/s12104-011-9348-8