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 D_9k 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.