Repetitive elements of the Mycoplasma hominis adhesin p50 can be differentiated by monoclonal antibodies.
The gene encoding p50, an adhesin of Mycoplasma hominis, was identified, cloned, and sequenced. Comparison of the derived amino acid sequence with the N-terminal amino acids sequenced by the Edman reaction of the native protein revealed that p50 is expressed as a 467-amino-acid precursor. Posttranslational modification leads to a 441-amino-acid lipoprotein with an extended, predominantly helical structure and a leucine zipper. Computer analysis of the amino acid sequence identified a threefold-repetitive sequence motif comprising approximately three-quarters of the total protein. Different regions of the p50 polypeptide chain were expressed in Escherichia coli. Western blot (immunoblot) analysis of the E. coli lysates revealed that the epitopes of four p50-specific monoclonal antibodies were localized in the middle and C-terminal part of the protein. Epitope mapping by exonuclease III digestion showed that all of the four monoclonal antibodies bound within the same region of the threefold-repetitive amino acid sequence motif. The repeats, which were highly homologous but not identical in structure, could be differentiated by the monoclonal antibodies.
Henrich
B
B
Institute for Medical Microbiology and Virology and Center for Biological and Medical Research, Heinrich-Heine-University, Duesseldorf, Germany.
Kitzerow
A
A
Feldmann
R C
RC
Schaal
H
H
Hadding
U
U
D018829:Adhesins, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D018604:Epitope Mapping; D008969:Molecular Sequence Data; D019535:Mycoplasma hominis / Q000737:chemistry*; D014158:Transcription, Genetic
D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal