Insight into the interaction of metal ions with TroA from Streptococcus suis

PLoS One. 2011;6(5):e19510. doi: 10.1371/journal.pone.0019510. Epub 2011 May 18.

Abstract

Background: The scavenging ability of sufficient divalent metal ions is pivotal for pathogenic bacteria to survive in the host. ATP-binding cassette (ABC)-type metal transporters provide a considerable amount of different transition metals for bacterial growth. TroA is a substrate binding protein for uptake of multiple metal ions. However, the function and structure of the TroA homologue from the epidemic Streptococcus suis isolates (SsTroA) have not been characterized.

Methodology/principal findings: Here we determined the crystal structure of SsTroA from a highly pathogenic streptococcal toxic shock syndrome (STSS)-causing Streptococcus suis in complex with zinc. Inductively coupled plasma mass spectrometry (ICP-MS) analysis revealed that apo-SsTroA binds Zn(2+) and Mn(2+). Both metals bind to SsTroA with nanomolar affinity and stabilize the protein against thermal unfolding. Zn(2+) and Mn(2+) induce distinct conformational changes in SsTroA compared with the apo form as confirmed by both circular dichroism (CD) and nuclear magnetic resonance (NMR) spectra. NMR data also revealed that Zn(2+)/Mn(2+) bind to SsTroA in either the same site or an adjacent region. Finally, we found that the folding of the metal-bound protein is more compact than the corresponding apoprotein.

Conclusions/significance: Our findings reveal a mechanism for uptake of metal ions in S. suis and this mechanism provides a reasonable explanation as to how SsTroA operates in metal transport.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anilino Naphthalenesulfonates / metabolism
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism*
  • Binding Sites
  • Calorimetry
  • Circular Dichroism
  • Cloning, Molecular
  • Computational Biology
  • Crystallography, X-Ray
  • Electron Spin Resonance Spectroscopy
  • Fluorescence
  • Ions
  • Magnetic Resonance Spectroscopy
  • Manganese / metabolism
  • Metals / metabolism*
  • Models, Biological
  • Molecular Sequence Data
  • Protein Binding
  • Protein Conformation
  • Protein Stability
  • Sequence Alignment
  • Streptococcus suis / metabolism*
  • Temperature
  • Zinc / metabolism

Substances

  • Anilino Naphthalenesulfonates
  • Bacterial Proteins
  • Ions
  • Metals
  • Manganese
  • 1-anilino-8-naphthalenesulfonate
  • Zinc