Treponema pallidum in gel microdroplets

A novel strategy for investigation of treponemal molecular architecture

D. L. Cox, D. R. Akins, S. F. Porcella, M. V. Norgard, J. D. Radolf

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Controversy exists regarding the constituents and antigenic properties of the Treponema pallidum outer membrane; a major point of contention concerns the cellular location(s) of the spirochaete's lipoprotein immunogens. To address these issues and circumvent problems associated with prior efforts to localize treponemal surface antigens, we developed a novel strategy for investigating T. pallidum molecular architecture. Virulent treponemes were encapsulated in porous agarose beads (gel microdroplets) and then probed in the presence or absence of Triton X-100. Intact, encapsulated treponemes were not labelled by monospecific antisera directed against four major T. pallidum lipoproteins or a candidate T. pallidum outer membrane protein (TpN50) with C-terminal sequence homology to Escherichia coli OmpA or by human or rabbit syphilitic serum. Each of these immunologic reagents, however, labelled encapsulated treponemes co-incubated with detergent. In contrast, antibodies generated against isolated T. pallidum outer membranes labelled intact organisms and the pattern of fluorescence was consistent with the distribution of rare outer membrane proteins visualized by freeze-fracture electron microscopy. In addition to providing strong evidence that the protein portions of treponemal lipoproteins are located within the periplasmic space, these studies have extended our understanding of the topographical relationships among T. paliidum cell envelope constituents. They also demonstrate the feasibility of generating antibodies against rare outer membrane proteins and detecting them on the surfaces of virulent treponemes.

Original languageEnglish (US)
Pages (from-to)1151-1164
Number of pages14
JournalMolecular Microbiology
Volume15
Issue number6
StatePublished - 1995

Fingerprint

Treponema pallidum
Gels
Lipoproteins
Membrane Proteins
Periplasm
Spirochaetales
Membranes
Antibodies
Octoxynol
Surface Antigens
Sequence Homology
Detergents
Sepharose
Immune Sera
Electron Microscopy
Fluorescence
Escherichia coli
Rabbits
Serum
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Treponema pallidum in gel microdroplets : A novel strategy for investigation of treponemal molecular architecture. / Cox, D. L.; Akins, D. R.; Porcella, S. F.; Norgard, M. V.; Radolf, J. D.

In: Molecular Microbiology, Vol. 15, No. 6, 1995, p. 1151-1164.

Research output: Contribution to journalArticle

Cox, D. L. ; Akins, D. R. ; Porcella, S. F. ; Norgard, M. V. ; Radolf, J. D. / Treponema pallidum in gel microdroplets : A novel strategy for investigation of treponemal molecular architecture. In: Molecular Microbiology. 1995 ; Vol. 15, No. 6. pp. 1151-1164.
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