Epitope mapping of the variable repetitive region with the MB antigen of Ureaplasma urealyticum.
One of the major surface structures of Ureaplasma urealyticum recognized by antibodies of patients during infection is the MB antigen. Previously, we showed by Western blot (immunoblot) analysis that any one of the anti-MB monoclonal antibodies (MAbs) 3B1.5, 5B1.1, and 10C6.6 could block the binding of patient antibodies to MB. Subsequent DNA sequencing revealed that a unique six-amino-acid direct tandem repeat region composed the carboxy two-thirds of this antigen. In the present study, using antibody-reactive peptide scanning of this repeat region, we demonstrated that the amino acids defining the epitopes for MAbs 3B1.5 5B1.1 and 10C6.6 are EQP, GK, and KEQPA, respectively. Peptide scanning analysis of an infected patient's serum antibody response showed that the dominant epitope was defined by the sequence PAGK. Mapping of these continuous epitopes revealed overlap between all MAb and patient polyclonal antibody binding sites, thus explaining the ability of a single MAb to apparently block all polyclonal antibody binding sites. We also show that a single amino acid difference in the sequence of the repeats of serovars 3 and 14 accounts for the lack of reactivity with serovar 14 of two of the serovar 3-specific MAbs. Finally, the data demonstrate the need to obtain the sequences of the mba genes of all serovars before an effective serovar-specific antibody detection method can be developed.
Zheng
X
X
Department of Microbiology, University of Alabama, Birmingham Schools of Medicine and Dentistry 35294, USA.
Lau
K
K
Frazier
M
M
Cassell
G H
GH
Watson
H L
HL
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry*; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid / Q000276:immunology*; D016990:Ureaplasma urealyticum / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; C089269:multiple-banded antigen, Ureaplasma