The 34-kilodalton membrane immunogen of Treponema pallidum is a lipoprotein.
Treponema pallidum subsp. pallidum and Escherichia coli incorporated exogenous [3H]palmitate into the 34-kilodalton (kDa) pathogen-specific antigen of T. pallidum. Radiolabeled fatty acid remained associated with the protein upon immunoprecipitation and after boiling in sodium dodecyl sulfate, acetone precipitation, and extensive extractions in organic solvents, suggesting that the fatty acid was covalently bound to the protein. Detection of [3H]palmitate after alkaline and acid hydrolyses confirmed the identity of the incorporated label. Globomycin inhibited maturation of the recombinant 34-kDa antigen, suggesting that E. coli uses the lipoprotein-specific signal peptidase II to process the treponemal antigen. Globomycin also inhibited processing of the 34-kDa antigen, as well as the 44.5- and 15-kDa antigens, in T. pallidum, implying that T. pallidum also possesses the lipoprotein export pathway common to both gram-negative and gram-positive bacteria. Ethanol inhibited processing of the 34-kDa antigen in minicells, suggesting that the 34-kDa antigen normally is translocated through the cytoplasmic membrane. Comparison of the Triton X-114 phase partitioning behavior of the 34-kDa antigen produced either by minicells or by a cell-free translation system indicated that the covalent attachment of fatty acid conferred hydrophobic biochemical properties to the 34-kDa antigen, consistent with the hypothesis that the attached lipid anchors the 34-kDa antigen into the membrane.
Department of Microbiology, University of Texas, Southwestern Medical Center, Dallas 75235.
M A
MA
Swancutt
J D
JD
Radolf
M V
MV
Norgard
D000215:Acylation; D000900:Anti-Bacterial Agents; D000942:Antigens, Bacterial / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D000431:Ethanol / Q000494:pharmacology; D005227:Fatty Acids / Q000494:pharmacology; D008074:Lipoproteins / Q000032:analysis*; D017830:Octoxynol; D010455:Peptides / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011233:Precipitin Tests; D014210:Treponema pallidum / Q000276:immunology*
D000900:Anti-Bacterial Agents; D000942:Antigens, Bacterial; D005227:Fatty Acids; D008074:Lipoproteins; D010455:Peptides; D000431:Ethanol; D011092:Polyethylene Glycols; C017339:globomycin; D017830:Octoxynol; C010615:Nonidet P-40