MOBILITY AND SYMMETRY IN THE FC AND PFC' FRAGMENTS AS PROBED BY H-1-NMR

The 270 MHz 1H NMR spectra of rabbit Fc and pFc′ fragments appear well resolved when compared to the spectra of other proteins of similar molecular weight. This is interpreted as evidence for substantial segmental flexibility in these fragments. This mobility can be monitored using two special NMR pulse sequences which exploit either the multiplet structure (spin coupling) or the differential linewidths of resonances. Several aromatic residues with mobility independent of the rest of the protein could be detected by these techniques. The titration behaviour of the histidine residues of pFc′ indicates that this fragment possesses a longitudinal C-2 symmetry axis. Of the three pairs of histidines in pFc′, two titrate with pKa values of 6.8 and 7.3, while the remaining pair remains protonated over the pH range 5.2-8.2 suggesting that these residues are not readily accessible to the solvent. The observation of five pairs of histidine C-2 proton resonances in the Fc indicates that the symmetry axis is retained in this larger fragment. Cleavage of the inter-heavy chain disulphide bond in the Fc fragment has little effect on its NMR spectrum. We find from this that the reduction in complement binding efficiency associated with this cleavage is not a direct consequence of a concomitant large structural change in the Fc fragment. © 1979.