Surface immunolabeling and consensus computational framework to identify candidate rare outer membrane proteins of Treponema pallidum

Infect Immun. 2010 Dec;78(12):5178-94. doi: 10.1128/IAI.00834-10. Epub 2010 Sep 27.


Treponema pallidum reacts poorly with the antibodies present in rabbit and human syphilitic sera, a property attributed to the paucity of proteins in its outer membrane. To better understand the basis for the syphilis spirochete's "stealth pathogenicity," we used a dual-label, 3-step amplified assay in which treponemes encapsulated in gel microdroplets were probed with syphilitic sera in parallel with anti-FlaA antibodies. A small (approximately 5 to 10%) but reproducible fraction of intact treponemes bound IgG and/or IgM antibodies. Three lines of evidence supported the notion that the surface antigens were likely β-barrel-forming outer membrane proteins (OMPs): (i) surface labeling with anti-lipoidal (VDRL) antibodies was not observed, (ii) immunoblot analysis confirmed prior results showing that T. pallidum glycolipids are not immunoreactive, and (iii) labeling of intact organisms was not appreciably affected by proteinase K (PK) treatment. With this method, we also demonstrate that TprK (TP0897), an extensively studied candidate OMP, and TP0136, a lipoprotein recently reported to be surface exposed, are both periplasmic. Consistent with the immunolabeling studies, TprK was also found to lack amphiphilicity, a characteristic property of β-barrel-forming proteins. Using a consensus computational framework that combined subcellular localization and β-barrel structural prediction tools, we generated ranked groups of candidate rare OMPs, the predicted T. pallidum outer membrane proteome (OMPeome), which we postulate includes the surface-exposed molecules detected by our enhanced gel microdroplet assay. In addition to underscoring the syphilis spirochete's remarkably poor surface antigenicity, our findings help to explain the complex and shifting balance between pathogen and host defenses that characterizes syphilitic infection.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Surface / immunology
  • Bacterial Outer Membrane Proteins / immunology*
  • Bacterial Outer Membrane Proteins / metabolism
  • Cloning, Molecular
  • Computational Biology / methods
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique / methods
  • Immunoblotting
  • Male
  • Rabbits
  • Syphilis / immunology
  • Syphilis / microbiology
  • Treponema pallidum / immunology*
  • Treponema pallidum / metabolism


  • Antigens, Surface
  • Bacterial Outer Membrane Proteins