Physicochemical evidence that Treponema pallidum TroA is a zinc-containing metalloprotein that lacks porin-like structure

J Bacteriol. 1999 Jul;181(14):4420-3. doi: 10.1128/JB.181.14.4420-4423.1999.

Abstract

Although TroA (Tromp1) was initially reported to be a Treponema pallidum outer membrane protein with porin-like properties, subsequent studies have suggested that it actually is a periplasmic substrate-binding protein involved in the transport of metals across the treponemal cytoplasmic membrane. Here we conducted additional physicochemical studies to address the divergent viewpoints concerning this protein. Triton X-114 phase partitioning of recombinant TroA constructs with or without a signal sequence corroborated our prior contention that the native protein's amphiphilic behavior is due to its uncleaved leader peptide. Whereas typical porins are trimers with extensive beta-barrel structure, size exclusion chromatography and circular dichroism spectroscopy revealed that TroA was a monomer and predominantly alpha-helical. Neutron activation, atomic absorption spectroscopy, and anomalous X-ray scattering all demonstrated that TroA binds zinc in a 1:1 molar stoichiometric ratio. TroA does not appear to possess structural features consistent with those of bacterial porins.

Publication types

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

MeSH terms

  • Bacterial Proteins
  • Circular Dichroism
  • Metalloproteins / chemistry*
  • Metalloproteins / genetics
  • Metalloproteins / metabolism
  • Octoxynol
  • Polyethylene Glycols
  • Porins / chemistry*
  • Porins / genetics
  • Porins / metabolism
  • Spectrometry, X-Ray Emission
  • Spectrophotometry, Atomic
  • Treponema pallidum / chemistry*
  • Treponema pallidum / genetics
  • Treponema pallidum / metabolism
  • Zinc / chemistry*
  • Zinc / metabolism*

Substances

  • Bacterial Proteins
  • Metalloproteins
  • Porins
  • Tromp1 protein, Treponema pallidum
  • Polyethylene Glycols
  • Octoxynol
  • Nonidet P-40
  • Zinc