Proteins electron paramagnetic resonance

The crystallographic structure of rubredoxin from Clostridium pasteurianum at 2.5 A, a resolution sufficient to reveal the sequence of several of the bulky amino acid side chains, shows the iron coordinated to two pairs of cysteine residues located rather near the termini of the polypeptide chain (Fig. 1). A related rubredoxin, with a three times larger molecular weight, from Pseudomonas oleovorans is believed to bind iron in a similar fashion. This conclusion is based on physical probes, especially electron paramagnetic resonance spectroscopy, all of which indicate that the iron is in each case situated in a highly similar environment however, the proteins display some specificity in catalytic function. 

Yang, A.-S. and B. J. Gaffney (1987). Determination of relative spin concentration in some high-spin ferric proteins using E/D distribution in electron paramagnetic resonance simulations. Biophys. J. 51 55-67. 

Davydov, R., Kuprin, S. Graslund, A., and Ehrenberg, A. 1994. Electron paramagnetic resonance study of the mixed-valent diiron center in Escherichia coli ribonucleotide reductase produced by reduction of radical-free protein R2 at 77 K. Journal of the American Chemical Society 116 11120-11128. 

Venable, J.H. 1967. Electron paramagnetic resonance spectroscopy of protein single crystals II. Computational methods. In Magnetic Resonance in Biological Systems, eds. A. Ehrenberg, B.G. Malmstrom and T. Vanngard Elmsford. New York Pergamon Press, 373-381. 

J. Zuclich, Triplet-state electron paramagnetic resonance of the aromatic amino acids and proteins, J. Chem. Phys. 52, 3586-3591 (1970). 

Based on our current understanding of ribosomal protein synthesis, several strategies have been developed to incorporate amino acids other than the 20 standard proteinogenic amino acids into a peptide using the ribosomal machinery . This allows for the design of peptides with novel properties. On the one hand, such a system can be used to synthesize nonstandard peptides that are important pharmaceuticals. In nature, such peptides are produced by nonribosomal peptide synthetases, which operate in complex pathways. On the other hand, non-natural residues are a useful tool in biochemistry and biophysics to study proteins. For example, incorporation of non-natural residues by the ribosome allows for site-specific labeling of proteins with spin labels for electron paramagnetic resonance spectroscopy, with... 

Selected entries from Methods in Enzymology [vol, page(s)] Electron paramagnetic resonance [effect on line width, 246, 596-598 motional narrowing spin label spectra, 246, 595-598 slow motion spin label spectra, 246, 598-601] helix-forming peptides, 246, 602-605 proteins, 246, 595 Stokes-Einstein relationship, 246, 594-595 temperature dependence, 246, 602, 604. 

The FeMo cofactor (or M center) in the MoFe-proteinis in the native paramagnetic M state. Reduction of the MoFe-protein by the Fe-protein results in the reduction of FeMo-co from the M state to the M state at a potential estimated to be less than —0.465 V (NHE). The electron paramagnetic resonance (EPR) silent M state is only transiently produced during catalysis, and relaxes to the M state when catalysis stops. The intimate consequences of the M state reduction are not precisely known. A more oxidized diamagnetic state may also be generated (M ) at —0.042 V but its biological relevance is unclear [9]. 

Henry, Y., Ducrocq, C., Drapier, J. C., Servent, D., Pellat, C., and Guissani, A. (1991). Nitric oxide, a biological effector. Electron paramagnetic resonance detection of nitrosyl-iron-protein complexes in whole cells. Eur. Biophys. J. 20, 1-15. 

Pellat, C., Henry, Y., and Drapier, j. C. (1990). IFN-gamma-activated macrophages Detection by electron paramagnetic resonance of complexes between L-arginine-derived nitric oxide and nonheme iron proteins. Biochem. Biophys. Res. Commun. 166, 119-125. 

The discussion of activity from X-ray data in conjunction with kinetic data is also difficult because, apart from considerations of dynamics, these techniques do not provide the essential knowledge about the energy states of given atoms or groups. It is necessary to inspect the electronic structure of at least certain regions of the protein. Methods exist for this inspection, and these include electron paramagnetic resonance, ultraviolet, circular dichroism, Raman and Mossbauer spectroscopies. The full understanding of activity can only come when the information derived from all available methods is assimilated and rationalized. 

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