title: Heme binding in the NEAT domains of IsdA and IsdC of Staphylococcus aureus.
abstract: Absorption, magnetic circular dichroism (MCD), and electrospray mass spectral (ESI-MS) data are reported for the heme binding NEAr iron Transporter (NEAT) domains of IsdA and IsdC, two proteins involved in heme scavenging by Staphylococcus aureus. The mass spectrometry data show that the NEAT domains are globular in structure and efficiently bind a single heme molecule. In this work, the IsdA NEAT domain is referred to as NEAT-A, the IsdC NEAT domain is referred to as NEAT-C, heme-free NEAT-C is NEAT-A and NEAT-C are inaccessible to small anionic ligands. Reduction of the high-spin Fe(III) heme iron to 5-coordinate high-spin Fe(II) in NEAT-A results in coordination by histidine and opens access, allowing for CO axial ligation, yielding 6-coordinate low-spin Fe(II) heme. In contrast, reduction of the high-spin Fe(III) heme iron to 5-coordinate high-spin Fe(II) in NEAT-C results in loss of the heme from the binding site of the protein due to the absence of a proximal histidine. The absorption and MCD data for NEAT-A closely match those previously reported for the whole IsdA protein, providing evidence that heme binding is primarily a property of the NEAT domain.
authors.affiliation: Department of Chemistry, University of Western Ontario, London, Ontario, Canada N6A 5B7.
authors.forename: Mark
authors.initials: M
authors.lastname: Pluym
authors.forename: Naomi
authors.initials: N
authors.lastname: Muryoi
authors.forename: David E
authors.initials: DE
authors.lastname: Heinrichs
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Stillman
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002942:Circular Dichroism; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
chemical_list: D000942:Antigens, Bacterial; D002352:Carrier Proteins; C512791:IsdA protein, Staphylococcus aureus; C488727:IsdC protein, Staphylococcus aureus; D006418:Heme