Anisotropy proteins

Tjandra N, Szabo A and Bax A 1996 Protein backbone dynamics and N-15 chemical shift anisotropy from quantitative measurement of relaxation interference effected. Am. Chem. Soc. 118 6986-91... 

Tjandra N and Bax A 1997 Solution NMR measurement of amide proton chemical shift anisotropy in N-15-enriched proteins. Correlation with hydrogen bond length J. Am. Chem. Soc. 119 8076-82... 

DIffey W M, Homoelle B J, Edington M D and Beck W F 1998 Excited-state vibrational coherence and anisotropy decay In the bacterlochlorophyll a dimer protein B820 J. Phys. Chem. B 102 2776-86... 

With these assignments at hand the analysis of the hyperfine shifts became possible. An Fe(III) in tetrahedral structures of iron-sulfur proteins has a high-spin electronic structure, with negligible magnetic anisotropy. The hyperfine shifts of the protons influenced by the Fe(III) are essentially Fermi contact in origin 21, 22). An Fe(II), on the other hand, has four unpaired electrons and there may be some magnetic anisotropy, giving rise to pseudo-contact shifts. In addition, there is a quintet state at a few hundred cm which may complicate the analysis of hyperfine shifts, but the main contribution to hyperfine shifts is still from the contact shifts 21, 22). 

Despite its weakness, the anisotropy of the g tensor of iron-sulfur centers can be used to determine the orientation of these centers or that of the accommodating polypeptide in relation to a more complex system such as a membrane-bound complex. For this purpose, the EPR study has to be carried out on either partially or fully oriented systems (oriented membranes or monocrystals, respectively). Lastly, the sensitivity of the EPR spectra of iron-sulfur centers to structural changes can be utilized to monitor the conformational changes induced in the protein by different factors, such as the pH and the ionic strength of the solvent or the binding of substrates and inhibitors. We return to the latter point in Section IV. 

The anisotropy in the physicochemical surface properties and differences in the surface topography of S-layer lattices allowed the determination of the orientation of the monolayers with respect to different surfaces and interfaces. Since in S-layers used for crystallization studies the outer surface is more hydrophobic than the inner one, the protein lattices were generally oriented with their outer face against the air-water interface [120,121]. Crystallization studies with the S-layer protein fromB. coagulansE i Nl at different lipid monolayers [122] revealed that the S-layer lattice is attached to lipid monolay-... 

Borst JW, Flink MA, van Hoek A, Visser AJWG (2005) Effects of refractive index and viscosity on fluorescence and anisotropy decays of enhanced cyan and yellow fluorescent proteins. JFluoresc 15 153-160... 

Shi X, Basran J, Seward FIE, Childs W, Bagshaw CR, Boxer SG (2007) Anomalous negative fluorescence anisotropy in yellow fluorescent protein (YFP 10C) quantitative analysis of FRET in YFP dimers. Biochemistry 46 14403-14417... 

Squire, A., Verveer, P. J., Pocks, O. and Bastiaens, P. I. H. (2004). Red-edge anisotropy microscopy enables dynamic imaging of homo-FRET between green fluorescent proteins in cells. J. Struct. Biol. 147, 62-9. 

Volkmer, A., Subramaniam, V., Birch, D. J. and Jovin, T. M. (2000). One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins. Biophys. J. 78, 1589-98. 

The intensity expression in Equation 6.5 requires all three g-values to be known. Sometimes not all g-values can be measured experimentally, and they have to be estimated on theoretical grounds. For example, the Fe(III) spectra of low-spin hemo-proteins frequently exhibit very pronounced g-anisotropy to the extent that two of the three g-values are either at helds beyond the maximum of the magnet and/or are associated with features inhomogeneously broadened beyond detection. With only the highest g-value determined the theoretical boundary condition for low-spin d5 systems with 3 < gz < 4... 

The versatility of luminescence goes beyond intensity-, wavelength- and kinetic-based measurements. Fluorescence polarization (or anisotropy) is an additional parameter still largely unexplored for optical sensing yet widely used in Biochemistry to study the interaction of proteins, the microfluidity of cell membranes and in fluorescence immunoassays. Although only a few optosensors based on luminescence polarization measurements can be found in the literature, elegant devices have recently been reported to measure chemical parameters such as pFI or O2 even with the bare eye41. 

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