Isolation and characterization of a novel IgD-binding protein from Moraxella catarrhalis.
A novel surface protein of the bacterial species Moraxella catarrhalis that displays a high affinity for IgD (MID) was solubilized in Empigen and isolated by ion exchange chromatography and gel filtration. The apparent molecular mass of monomeric MID was estimated to approximately 200 kDa by SDS-PAGE. The mid gene was cloned and expressed in Escherichia coli. The complete mid nucleotide gene sequence was determined, and the deduced amino acid sequence consists of 2123 residues. The sequence of MID has no similarity to other Ig-binding proteins and differs from all previously described outer membrane proteins of M. catarrhalis. MID was found to exhibit unique Ig-binding properties. Thus, in ELISA, dot blots, and Western blots, MID bound two purified IgD myeloma proteins, four IgD myeloma sera, and finally one IgD standard serum. No binding of MID was detected to IgG, IgM, IgA, or IgE myeloma proteins. MID also bound to the surface-expressed B cell receptor IgD, but not to other membrane molecules on human PBLs. This novel Ig-binding reagent promises to be of theoretical and practical interest in immunological research.
Forsgren
A
A
Department of Medical Microbiology, Malmö University Hospital, Lund University, Malmö, Sweden.
Brant
M
M
Möllenkvist
A
A
Muyombwe
A
A
Janson
H
H
Woin
N
N
Riesbeck
K
K
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology; D005822:Genetic Vectors / Q000276:immunology; D006801:Humans; D007072:Immunoglobulin D / Q000378:metabolism*; D008969:Molecular Sequence Data; D001936:Moraxella catarrhalis / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D020539:Sequence Analysis, Protein
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; C439616:IgD-binding protein MID, Moraxella catarrhalis; D007072:Immunoglobulin D