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 15N-labeled mutant of calbindin D9k loaded with either Tm3+ or Tb3+, reduced R1 and R2 relaxation rates of backbone 15N 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.Financial support by the Australian Research Council is gratefully acknowledged