Antigenic interrelationship between endoflagella of Treponema phagedenis biotype Reiter and Treponema pallidum (Nichols): molecular characterization of endoflagellar proteins

Abstract

Purified endoflagella from nonpathogenic Treponema phagedenis bioptye Reiter were characterized biochemically and compared antigenically with the endoflageilar proteins of Treponema pallidum. T. phagedenis biotype Reiter endoflagella were dissociated into constituent polypeptides by incubation under conditions which disrupt noncovalent bonds. Chymotrypsin peptide maps of T. phagedenis biotype Reiter endoflageliar proteins revealed that the 37and 33-kilodalton (kDa) major components shared significant homology with the 27and 30-kDa minor components, respectively. The peptide maps also suggested that the major components shared a lesser degree of structural similarity with each other. These relationships were confirmed by Western blots of T. phagedenis biotype Reiter endoflagellar proteins employing antibodies that were purified against the individual endoflagellar polypeptides. Western blots of T. pallidum with the purified antibodies also demonstrated strong cross-reactivity between the T. phagedenis biotype Reiter endoflageliar proteins and T. pallidum proteins of identical or similar molecular weights. A unique Western blotting technique that we called epitope bridging was used to determine that the 37-kDa subunit contains most of the external epitopes on T. phagedenis biotype Reiter endoflagella. Immunoelectron microscopy with human syphilitic serum and rabbit T. phagedenis biotype Reiter endoflagellar antiserum confirmed the presence of cross-reactive epitopes on the surface of intact T. phagedenis and T. paUidum endoflagella.

Common and specific antigens of several treponemes detected by polyclonal antisera against major cellular proteins

“…Many investigators have described axial fiagellar polypeptides with similar molecular sizes (8,13,14,19,20). T.patltdutn strain Nichols and 77 phagedenis have an additional flagellar protein with an approximate molecular weight of 38 kDa (2,13,17,19,20). The two ATCC strains of 77 dentieola possessed the 34 kDa P3 and 38 kDa PI 1 axial flagellar proteins.…”

“…Discontinuous SDS-PAGE was conducted on the basis of the method of Laemmli as reported previously (II). A mixture of each bacterial suspension (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) fig of protein per lane) and solubilization buffer was heated at 100 C for 5 min and applied to 1-or 2-mmthick SDS-10% acrylamide gels. A total of 13 polypeptides were selected on the basis of individual quantity, clear separation from neighboring protein bands and exclusion of the antigens with apparently similar molecular sizes.…”

Cloning, sequencing, and expression of two class B endoflagellar genes of Treponema pallidum subsp. pallidum encoding the 34.5- and 31.0-kilodalton proteins

“…Recent studies performed in our laboratory have demonstrated the presence of T. pallidum specific antigenic determinants on the 37-, 34.5-, 33-, and 31-kDa endoflagellar polypeptides (3). In view of the demonstrated cross-reactivity that exists among the endoflagella of T. pallidum, host indigenous nonpathogenic treponemes (3,4,13,24,29), and other pathogenic spirochetes, including Borrelia burgdorferi (18; Lovett et al, unpublished data), and Leptospira interrogans (Lovett et al, unpublished data), these unique T. pallidum endoflagellar epitopes could well provide a more specific and sensitive serodiagnostic antigen for the detection of syphilis.…”

“…Endoflagella are major protein and antigenic constituents of T. pallidum which are recognized by antibody early in and throughout the course of the disease (11,12,26,27). Serodiagnostic studies in which workers have used endoflagella prepared from Treponema phagedenis biotype Reiter, whose endoflagella share considerable antigenic cross-reactivity with the endoflagella of T. pallidum (3,4,13,24,29), have shown similar degrees of sensitivity and specificity compared with the standard T. pallidum serodiagnostic tests currently in use (27). Recent studies performed in our laboratory have demonstrated the presence of T. pallidum specific antigenic determinants on the 37-, 34.5-, 33-, and 31-kDa endoflagellar polypeptides (3).…”

Antigenic Behaviors of Two Axial Flagellar Proteins Detected in Treponema denticola

“…Purified axial flagella of T. pallidum Nichols and T. phagedenis biotype Reiter consist of at least two proteins with approximate molecular sizes of 33 kDa to 34 kDa and 37 kDa to 39 kDa, although their reported molecular sizes are not coincident among investigators (10,11). Limberger and Charon also reported a 34-kDa common antigen in several treponemes (7).…”