Determination of the electron relaxation rates in paramagnetic metal complexes: applicability of available NMR methods.

Four different approaches for determining the electron relaxation rates in paramagnetic metallo-proteins are investigated, using a paramagnetic Ni2+ complex of a protein as an example. All four approaches rely on the determination of the longitudinal paramagnetic relaxation enhancements, R1p, of the 1H nuclei and the backbone 15N nuclei. Three of the methods utilize the field dependence of the R1p rates. It is found that the applicability of each of these methods depends on whether the fast-motion condition, omegaS2tau2<<1, applies to the electron relaxation, omegaS being the Larmor frequency of the electron spin S and tau the correlation time of the electron relaxation. If the fast-motion condition is fulfilled, the electron relaxation rate can be obtained from the ratio of the R1p rates of one or more protons at two magnetic field strengths (method A). On the other hand, if the fast-motion condition does not apply, more elaborate methods must be used that, in general, require a determination of the R1p rates over a larger range of magnetic field strengths (method C). However, in the case of paramagnetic metal ions with relatively slow electron relaxation rates only two magnetic field strengths suffice, if the R1p rates of a hetero nucleus are included in the analysis (method B). In the fourth method (method D), the electron relaxation is estimated as a parameter in a structure calculation, using distance constraints derived from proton R1p rates at only one magnetic field strength. In general, only methods B and C give unambiguous electron relaxation rates.