Elsevier

Methods in Enzymology

Volume 339, 2001, Pages 314-340
Methods in Enzymology

[15] - Paramagnetic Probes in Metalloproteins

https://doi.org/10.1016/S0076-6879(01)39320-5Get rights and content

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Introduction: Solving Solution Structures of Paramagnetic Proteins

At the beginning of the 1990s, when obtaining solution structures of bio-molecules by nuclear magnetic resonance (NMR) was already a well-established technique1, 2 it was commonly believed that solution structures of macromolecules containing paramagnetic centers could not be solved.3 Indeed, the loss of information arising from the hyperfine interaction between nuclear spins and the unpaired electron spin was supposed to be (i) too severe and (ii) unavoidable.

Nevertheless, the first solution

Use of Nonconventional Constraints for Structure Calculations

Once it was demonstrated that many of the drawbacks due to the presence of a paramagnetic center can be overcome in favorable cases,5 the goal was to take a reverse approach. Hyperfine interaction may provide additional information that would simply not be there in diamagnetic systems. This chapter deals with the theoretical and methodological aspects related to the exploitation of the hyperfine interaction in terms of structural constraints.

After the first structure in solution was solved,4 we

Paramagnetic Probes and Orientation Constraints

In recent years there have been intense research efforts aimed at exploiting partial orientation of biomolecules in magnetic fields.126, 127, 128, 129 The goal is that of recovering detectability of dipolar interactions that are averaged zero in solution when the molecule reorients isotropically. The tendency for a molecule to partially orient in solution in a magnetic field B0 depends on its magnetic susceptibility anisotropy and on the square of the magnetic field.130 Among biomolecules,

Cross Correlation and Hyperfine Interaction

Cross-correlation phenomena contribute to both shifts and relaxation properties of NMR signals. Although they have been known for years, only recently has it been emphasized that these effects have great potential in providing detailed structural and dynamic information on biomolecules in solution153, 154 and in allowing important steps to be made in the study of large biological macromolecules.155

The interaction between nuclear spins and the static electron magnetic moment induced by the

Experimental Techniques

Next we will briefly discuss some tools that have been developed and used to study paramagnetic systems. Although there have been no earth-shaking new techniques from the experimental point of view, the need to obtain as much information as possible in the neighborhood of the metal ion has led several authors: (i) to develop building blocks specifically tailored for paramagnetic systems and to develop new approaches to identify extremely broad signals; and (ii) to better exploit the information

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