Conformational Studies of Partially Folded Protein by EPR and NMR Spin Labeling Methods: A Review

Abstract

Abstract:

Partially folded proteins, characterized as exhibiting secondary structure elements with loose or absent tertiary contacts, are important intermediates in physiological protein folding and under pathological conditions. Structural restraints derived from EPR and NMR spectroscopy can be used to test and build secondary, tertiary, and quaternary structural models as well as measure protein conformational changes in solution. Insertion of one or two cysteine residues into the protein backbone using molecular biology methods and the subsequent labeling of the cysteine residues with a paramagnetic spin label enables the techniques of both EPR and NMR to be used as the molecular spectroscopic ruler. Site-directed spin labeling electron paramagnetic resonance (SDSL-EPR) measures the dipolar interaction between pairs of paramagnetic spin labels to yield inter-nitroxide distances, while the method of paramagnetic relaxation enhancement (PRE) measured by NMR gives information on the distance between the spin labeling center and the nuclei. The inter-spin dipolar interactions at about 2.5 nm are measured using continuous wave (CW) EPR and NMR methods. As for any low-resolution distance methods, the positioning of the spin labels and the number of distance constraints to be measured are depends on the structural question being asked, thus a pattern approach for using distance sets to decipher structure mapping, including protein folds and conformational changes associated with biological activity, is essential.

Key words: EPR, NMR, partially folded protein, conformation, distance restraints

CLC Number:

O482.53

ZHU Lei, LIU Zhi-Jun. Conformational Studies of Partially Folded Protein by EPR and NMR Spin Labeling Methods: A Review[J]. Chinese Journal of Magnetic Resonance, 2011, 28(4): 523-533.

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