Difficulties in determining accurate molecular motion parameters from proton relaxation enhancement measurements as illustrated by the immunoglobulin G-Gd(III) system

Abstract

Longitudinal and traverse proton magnetic relaxation rates for water in the hydration sphere of Gd(III) bound to non-immune rabbit IgG (immunoglobulin G) have been determined over a wide range of frequencies (4-84 MHz) at constant temperature (19 degrees C) using pulsed nuclear magnetic resonance spectrometry. The rates have also been determined at temperatures between 0 and 40 degrees C for two frequencies (61 and 84 MHz). The rates were fitted to existing theory using a computer least-squares procedure. Further computer analysis was then carried out to determine the sensitivity of the best-fit error to variation in the variable parameters in the theoretical expressions used. These include the water co-ordination number (q) for which it was found large variations could occur (between approximately 2 and 8) for only small changes in the error of best-fit. It was concluded that a slow exchange contribution to the relaxation rates was important in deciding which parameters are poorly determined. A rotational correlation time (tau r) was obtained which suggested there might be considerable internal motion of the Fc region (C-terminal half of heavy-chain dimer) of the IgG molecule. However the possibility of large errors in this value prevented unequivocal conclusions being drawn.