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Characterization of the Outer Membrane of Treponema Pallidum Subsp. Pallidum by Binding Studies Using Antibodies, Complement, and Host Serum Proteins (open access)

Characterization of the Outer Membrane of Treponema Pallidum Subsp. Pallidum by Binding Studies Using Antibodies, Complement, and Host Serum Proteins

The major goal of this study was to achieve sustained cultivation of virulent T. pallidum in vitro. The putatuive binding of host proteins to the outer membrane (OM) of intact, virulent T. pallidum subsp. pallidum has been investigated. A major breakthrough was the development of a filtration assay, usinglow protein-binding membrane filters, for the measurement of substances bound to or incorporated into th eOM of T. pallidum. This avoided the conventional manipulations which can damage the fragile OM of T. pallidum. Using this filtration assay, studies on the binding of host serum proteins demonstrated that intact treponemes did not bind host proteins as previously reported. It also indicated that previous studies were probably performed with damaged by this research. The studies on the binding of polyclonal and monoclonal antibodies to intact and detergent treated treponemes provided evidence of the low level binding of antibody to intact treponemes which was greatly enhanced but the removal of the outer membrane with 0.1% Triton X. This research research corroborated that of others which suggests that the outer membrane of T. pallidum contains very little protein or surface exposed antigen.
Date: December 1989
Creator: Chang, Po-Hsun
System: The UNT Digital Library
Two-Dimensional Gel Electrophoresis of in Vivo and in Vitro Synthesized Proteins, Antigenic Proteins, and Cross-Reactive Antigens in Treponema Pallidum Subsp. Pallidum Nichols Strain and Treponema Phagedenis Biotype Reiter (open access)

Two-Dimensional Gel Electrophoresis of in Vivo and in Vitro Synthesized Proteins, Antigenic Proteins, and Cross-Reactive Antigens in Treponema Pallidum Subsp. Pallidum Nichols Strain and Treponema Phagedenis Biotype Reiter

Two-dimensional electrophoretic protein profiles of in vivo and in vitro propagated T.pallidum subsps. pallidum Nichols strain were analyzed and compared. This comparative analysis revealed two in vitro synthesized, cytoplasmic cylinder-associated polypeptides with molecular masses 29.5 and 34.7 kDa, pI 5.62, and one in vitro "lost" polypeptide with molecular mass 34.7 kDa, pI 5.34. integral membrane proteins of in vitro and in vivo propagated T. pallidum was identified by phase partitioning with the nonionic Triton X-114, and twelve outer membrane-associated, antigenic proteins were identified in western blots probed with pooled human secondary syphilitic sera. The solubilization of the outer membrane of T. pallidum with Triton X-114 were monitored by electron microscopy. Treatment of freshly harvested 35S labeled T. pallidum with 1% Triton X-114 resulted in solubilization of the outer membrane and reduction of the diameter of the treponemes from .14 +/- .02 micrometers to .095 +/- .003 micrometers. Examination of thin sections of untreated organisms showed integrity of outer and cytoplasmic membranes. In contrast, thin sections of Triton X-114-treated trponemes showed integrity of the cytoplasmic membrane but the loss of the outer membrane. The cytoplasmic cylinders generated by detergent treatment retained their periplasmic flagella, as judged by electron microscopy and immunoblotting. …
Date: May 1989
Creator: Sayahtaheri, Sousan
System: The UNT Digital Library