Proton-n.m.r. study of interaction of myelin basic protein with a monoclonal antibody
- 15 May 1985
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 228 (1), 61-68
- https://doi.org/10.1042/bj2280061
Abstract
Proton NMR at 400 MHz was applied to study the interactions of bovine or porcine myelin basic protein (b- or p-MBP) with a monoclonal antibody to human (h-) MBP. The antibody, an IgG that contains a sequential epitopic region, cross-reacts with b-MBP but not with p-MBP, the presumed epitope being identical in h- and b-MBP. NMR spectra were recorded from the Fab fragment of the antibody and for mixtures of Fab and MBP at various molar ratios. The NMR spectrum of MBP in the mixture consists mostly of well resolved peaks against a broad background due to the Fab. With b-MBP, but not p-MBP, specific interactions are observed at the residue tyrosine-135, which is part of the epitopic sequence. Other interactions occur between the Fab and both b- and p-MBP at residues distant from the epitopic region. Standard radioassay techniques were employed to calculate the binding constants of both basic proteins with the Ig. The binding constant, Kb, for IgG to column-immobilized b-MBP at 298 K is (0.95 .+-. 0.07) .times. 10- dm3/mol. The value of Kb decreases with the ionic strength of the medium, suggesting a coulombic interaction between antigen and antibody. NMR spectra were also measured for mixtures of the Fab fragment and peptides containing the epitopic site, with results in agreement with those for the whole protein.This publication has 15 references indexed in Scilit:
- Antigenic determinant recognized by a monoclonal antibody to human myelin basic proteinJournal of Neuroimmunology, 1983
- NMR studies of myelin basic proteinBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1983
- A computer program for predicting protein antigenic determinantsMolecular Immunology, 1983
- Carbon-13 nuclear magnetic resonance study of the binding of carbon-13-enriched tetra-L-alanine haptens to Fab' fragments of anti-poly(L-alanine) antibodiesBiochemistry, 1980
- [6] Preparation of Fab fragments from IgGs of different animal speciesMethods in Enzymology, 1980
- Thermodynamic analysis of ion effects on the binding and conformational equilibria of proteins and nucleic acids: the roles of ion association or release, screening, and ion effects on water activityQuarterly Reviews of Biophysics, 1978
- Structure of an antibody combining site by magnetic resonanceNature, 1977
- Continuous cultures of fused cells secreting antibody of predefined specificityNature, 1975
- Allergic encephalomyelitis. Oxidation and cleavage of the single tryptophan residue of the A1 protein from bovine and human myelin.1971
- Experimental allergic encephalomyelitisArchives of Biochemistry and Biophysics, 1969