title: Kinetics of heme transfer by the Shr NEAT domains of Group A Streptococcus.
abstract: The hemolytic Group A Streptococcus (GAS) is a notorious human pathogen. Shr protein of GAS participates in iron acquisition by obtaining heme from host hemoglobin and delivering it to the adjacent receptor on the surface, Shp. Heme is then conveyed to the SiaABC proteins for transport across the membrane. Using rapid kinetic studies, we investigated the role of the two heme binding NEAT modules of Shr. Stopped-flow analysis showed that holoNEAT1 quickly delivered heme to apoShp. HoloNEAT2 did not exhibit such activity; only little and slow transfer of heme from NEAT2 to apoShp was seen, suggesting that Shr NEAT domains have distinctive roles in heme transport. HoloNEAT1 also provided heme to apoNEAT2, by a fast and reversible process. To the best of our knowledge this is the first transfer observed between isolated NEAT domains of the same receptor. Sequence alignment revealed that Shr NEAT domains belong to two families of NEAT domains that are conserved in Shr orthologs from several species. Based on the heme transfer kinetics, we propose that Shr proteins modulate heme uptake according to heme availability by a mechanism where NEAT1 facilitates fast heme delivery to Shp, whereas NEAT2 serves as a temporary storage for heme on the bacterial surface.
authors.affiliation: Department of Biology, College of Arts and Sciences, Georgia State University, Atlanta, GA 30303, USA.
authors.forename: Mahamoudou
authors.initials: M
authors.lastname: Ouattara
authors.forename: Andrea
authors.initials: A
authors.lastname: Pennati
authors.forename: Darius J
authors.initials: DJ
authors.lastname: Devlin
authors.forename: Ya-Shu
authors.initials: YS
authors.lastname: Huang
authors.forename: Giovanni
authors.initials: G
authors.lastname: Gadda
authors.forename: Zehava
authors.initials: Z
authors.lastname: Eichenbaum
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D007700:Kinetics; D008706:Methemoglobin / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013290:Streptococcal Infections / Q000378:metabolism* / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000737:chemistry / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D006418:Heme; D008706:Methemoglobin
keywords: Gram-positive; Heme transfer; Kinetics; Methemoglobin; NEAT domain; Shr; Stopped-flow; Streptococcus pyogenes; Structure/function