Overproduction and purification of Treponema pallidum recombinant-DNA-derived proteins TmpA and TmpB and their potential use in serodiagnosis of syphilis.
We report the construction of expression plasmids carrying two Treponema pallidum genes encoding for the 42-kilodalton membrane protein TmpA (treponemal membrane protein A) and the 34-kilodalton membrane protein TmpB. Using the leftward promoter of bacteriophage lambda, which is controlled by a thermosensitive repressor, we obtained a high level of heat-inducible synthesis of TmpA and TmpB in Escherichia coli K-12. Both proteins were purified to near homogeneity, and the presence of antibodies to TmpA and TmpB in human sera was determined by an enzyme-linked immunosorbent assay. Whereas in all 44 serum samples from untreated patients in the secondary and early latent stages of syphilis, high levels of anti-TmpA antibodies were detected, only 34 serum samples contained anti-TmpB antibodies. As has been previously observed for TmpA, a correlation was found between the presence of anti-TmpB antibodies and anti-cardiolipin antibodies, suggesting that the level of antibodies to TmpB drops soon after successful antibiotic treatment. We concluded that, in contrast to TmpA, TmpB is not suitable for serodiagnostic purposes as a single antigen, because a significant fraction of sera from syphilitic patients was nonreactive with TmpB.
Schouls
L M
LM
Laboratory for Bacteriology, National Institute of Public Health and Environmental Hygiene, Bilthoven, The Netherlands.
Ijsselmuiden
O E
OE
Weel
J
J
van Embden
J D
JD
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D012698:Serologic Tests; D013587:Syphilis / Q000175:diagnosis* / Q000382:microbiology; D014210:Treponema pallidum / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins