Molecular Analysis of the Gene Encoding a Protein Component of the Eikenella Corrodens Adhesin Complex That Is Close to the Carbohydrate Recognition Domain

Gene. 1996 Nov 21;180(1-2):207-12. doi: 10.1016/s0378-1119(96)00639-7.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / genetics*
  • Adhesins, Bacterial / metabolism
  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology
  • Base Sequence
  • Blotting, Western
  • Cloning, Molecular
  • DNA, Bacterial
  • Eikenella corrodens / genetics*
  • Eikenella corrodens / immunology
  • Escherichia coli
  • Genes, Bacterial
  • Lectins / genetics
  • Lectins / immunology
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Restriction Mapping

Substances

  • Adhesins, Bacterial
  • Antibodies, Monoclonal
  • DNA, Bacterial
  • LecA protein, bacteria
  • Lectins
  • Recombinant Proteins

Associated data

  • GENBANK/D78153