Kinetics of heme transfer by the Shr NEAT domains of Group A Streptococcus

Arch Biochem Biophys. 2013 Oct 15;538(2):71-9. doi: 10.1016/j.abb.2013.08.009. Epub 2013 Aug 28.

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.

Keywords: Gram-positive; Heme transfer; Kinetics; Methemoglobin; NEAT domain; Shr; Stopped-flow; Streptococcus pyogenes; Structure/function.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Heme / metabolism*
  • Host-Pathogen Interactions*
  • Humans
  • Kinetics
  • Methemoglobin / metabolism
  • Protein Structure, Tertiary
  • Streptococcal Infections / metabolism*
  • Streptococcal Infections / microbiology
  • Streptococcus pyogenes / chemistry
  • Streptococcus pyogenes / physiology*

Substances

  • Bacterial Proteins
  • Heme
  • Methemoglobin