Molecular analysis of the gene encoding a protein component of the Eikenella corrodens adhesin complex that is close to the carbohydrate recognition domain.
A monoclonal antibody against a lectin-like substance (LS) of Eikenella corrodens (Ec) was used for screening the Ec DNA library. Three positive clones that carried an identical 12-kb segment were obtained. A 25-kDa protein, which specifically binds to the antibody, was overproduced in all of the Escherichia coli clones. Deletion analysis showed that the gene encoding the 25-kDa protein was located within a 1.2-kb segment. The nucleotide (nt) sequence of this segment contained an open reading frame encoding a protein of 24600 Da. We purified the 25-kDa protein from the cloned E. coli strain. The sequence of the first 10 amino acids(aa) from the N-terminus of the purified 25-kDa protein agreed with that deduced from the nt sequence. Since the monoclonal antibody used in this study inhibits the physiological activity of EcLS, we concluded that the 25-kDa protein is a component of the adhesin complex, which is located near the carbohydrate recognition domain of lectin in EcLS.
Azakami
H
H
Department of Conservative Dentistry, Tokushima University School of Dentistry, Japan. azakami@dent.dent.tokushima-u.ac.jp
Yumoto
H
H
Nakae
H
H
Matsuo
T
T
Ebisu
S
S
D018829:Adhesins, Bacterial / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D016124:Eikenella corrodens / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli; D005798:Genes, Bacterial; D037102:Lectins / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D015183:Restriction Mapping
D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D004269:DNA, Bacterial; C102929:LecA protein, bacteria; D037102:Lectins; D011994:Recombinant Proteins