The immunoglobulin-binding Eib proteins from Escherichia coli are receptors for IgG Fc.
The immunoglobulin-binding proteins from Escherichia coli (Eibs) comprise a family of six proteins homologous to the Yersinia adhesin YadA. These proteins are postulated to bind to the Fc portion of immunoglobulin G (IgG) in a non-immune manner. However, a recent study [Ghumra, A., Pleass, R.J., 2007. Escherichia coli do not express Fc-receptors for human immunoglobulin G (IgG). Mol. Immunol. 44, 2144-2146] appeared to show that these proteins do not bind Fc and suggested that the binding seen in earlier studies is due to the polyclonal preparations used in the assays containing antibodies specific to epitopes in the Eib proteins. To resolve this matter, we produced purified, recombinant Eibs for the first time and investigated their binding to intact antibodies and Fc fragments by immunoblot and ELISA techniques. We were able to purify four members of the family, EibA, -C, -D and -F, and show conclusively that these bind IgG Fc. We were also able to block the binding of full-length antibody with IgG Fc, but not with IgG Fab. Binding to IgG Fab was not detectable by surface plasmon resonance, whereas the affinities of Eibs to IgG and IgG Fc were in the range of 50-200 nM. We further demonstrate that deglycosylating IgG Fc does not affect Eib binding. Our results show that the Eib proteins do indeed bind human IgG Fc and that IgG Fc receptors are present in E. coli.
Leo
Jack C
JC
Macromolecular Crystallography Group, Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland.
Goldman
Adrian
A
D018830:Adhesins, Escherichia coli / Q000378:metabolism / Q000502:physiology; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000502:physiology; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism* / Q000502:physiology*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology; D011485:Protein Binding; D011971:Receptors, Immunologic / Q000378:metabolism* / Q000502:physiology; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism
D018830:Adhesins, Escherichia coli; D002352:Carrier Proteins; C440641:EibA protein, E coli; C440640:EibF protein, E coli; C514972:EibG protein, E coli; D029968:Escherichia coli Proteins; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011971:Receptors, Immunologic; D011994:Recombinant Proteins