Using Paramagnetism to Slow Down Nuclear Relaxation in Protein NMR

Paramagnetic metal ions accelerate nuclear spin relaxation; this effect is widely used for distance measurement and called paramagnetic relaxation enhancement (PRE). Theoretical predictions established that, under special circumstances, it is also possible to achieve a reduction in nuclear relaxation rates (negative PRE). This situation would occur if the mechanism of nuclear relaxation in the diamagnetic state is counterbalanced by a paramagnetic relaxation mechanism caused by the metal ion. Here we report the, first experimental evidence for such a cross-correlation effect. Using a uniformly N-15-labeled mutant of calbindin D-9k loaded with either Tm3+ or Tb3+, reduced R-1 and R-2 relaxation rates of backbone N-15 spins were observed compared with the diamagnetic reference (the same protein loaded with Y3+). The effect arises from the compensation of the chemical shift anisotropy tensor by the anisotropic dipolar shielding generated by the unpaired electron spin.