Determination of the Structural Basis of Antibody Diversity Using NMR.

摘要

A number of interesting discoveries have been made in our recent study of the monoclonal anti-spin label antibody AN02. (a) Structure-Kinetics: A tyrosine residue, Tyr31L, takes on one of two possible conformations when dinitrophenyl hapten binds. When the hapten dissociates, this tyrosine rotates rapidly. (b) AN02 is an auto antibody whose dimerization involves Tyr 31L and is blocked by hapten. (c) The technique of 'spin-label titration' to measure spin-to-proton distances has been verified experimentally. This has required two definitive proton resonance signal assignments for tyrosine resonances in the combining site, and comparison with the crystallographic results (Leahy Fox, to be published.) (d) Chemical shift calculations using ring current shielding calculations have proven remarkably accurate and useful. In addition to the above, a number of single site mutations have been prepared, expressed, and NMR spectra taken. (JS)