Anchoring antibodies to membranes using a diphtheria toxin T domain-ZZ fusion protein as a pH sensitive membrane anchor.
We have constructed a fusion protein, T-ZZ, in which the IgG-Fc binding protein ZZ was fused to the C-terminus of the diphtheria toxin transmembrane domain (T domain). While soluble at neutral pH, T-ZZ retained the capacity of the T domain to bind to phospholipid membranes at acidic pH. Once anchored to the membrane, the ZZ part of the protein was capable of binding mouse monoclonal or rabbit polyclonal IgG. Our results show that the T-ZZ protein can function as a pH sensitive membrane anchor for the linkage of IgG to the membrane of lipid vesicles, adherent and non-adherent cells.
Nizard
P
P
Département d'Ingénierie et d'Etudes des Protéines (DIEP), DSV, CEA, CE Saclay, Gif-sur-Yvette, France.
Liger
D
D
Gaillard
C
C
Gillet
D
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects; D004167:Diphtheria Toxin / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000378:metabolism*; D008081:Liposomes / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D013205:Staphylococcal Protein A / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004167:Diphtheria Toxin; D007074:Immunoglobulin G; D008081:Liposomes; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A