Binding characteristics of IgA 16.4.12E, a monoclonal antibody with specificity for the nonreducing terminal epitope of alpha-(1----6)-dextrans. Comparisons between IgA hybridoma 16.4.12E and myeloma W3129.
IgA 16.4.12E is a murine monoclonal antibody obtained following immunization with isomaltohexose linked to keyhole limpet hemocyanin. We have studied its interaction with methyl alpha-D-glucopyranoside and its derivatives bearing deoxy or deoxyfluoro groups, and with the methyl alpha-glycosides of a series of isomalto-oligosaccharides, some bearing deoxy or deoxy-fluoro groups at selected positions. From the data it is concluded that the antibody binds optimally to 4 sequential glucopyranosyl residues and that the protein subsite possessing the major affinity binds the terminal, nonreducing glucosyl group of that antigenic epitope. All the hydroxyl groups of that terminal glucosyl group are involved in hydrogen bonding, some in a donating and some in an accepting capacity. In the last part of the paper we report the construction of a possible model of the antibody, derived from its known amino acid sequence and the known crystalline structures of two closely related antibodies. It shows a pronounced cavity in the general immunoglobulin combining area which is flanked by 2 solvent-exposed tryptophanyl residues. A model recently reported for anti-dextran IgA W3129 shows a similar cavity with one such residue. Guided by hydrogen bonds, experimentally deduced from the comparison of the affinities of variously derivatized ligands, we suggest a speculative fitting for the nonreducing terminus of the dextran antigen, in the respective cavities of both IgA 16.4.12E and W3129.
Nashed
E M
EM
National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
Perdomo
G R
GR
Padlan
E A
EA
Kovác
P
P
Matsuda
T
T
Kabat
E A
EA
Glaudemans
C P
CP
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001666:Binding Sites, Antibody; D002236:Carbohydrate Conformation; D003911:Dextrans / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D006860:Hydrogen Bonding; D007070:Immunoglobulin A / Q000276:immunology* / Q000378:metabolism; D008024:Ligands; D008757:Methylglucosides / Q000138:chemical synthesis; D008958:Models, Molecular; D009194:Myeloma Proteins / Q000276:immunology; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
D000911:Antibodies, Monoclonal; D003911:Dextrans; D000939:Epitopes; D007070:Immunoglobulin A; D008024:Ligands; D008757:Methylglucosides; D009194:Myeloma Proteins