Nanoparticles consisting of either a solid core of GdF3 or an 80/20 mixture of GdF3 and LaF3 have been prepared for use as NMR and MRI relaxation agents. To obtain high aqueous solubilities, the particles were coated with either citrate (cit) groups (in the case of GdF3 nanoparticles), giving the nanoparticle a negatively charged surface, or 2-aminoethyl phosphate (AEP) groups (in the case of GdF3/LaF3 = 80/20), giving the nanoparticle a positively charged surface at physiological pH. In the presence of the 80/20 GdF3/LaF3:AEP, the paramagnetic contribution to the water spin−lattice relaxation rate was observed to be 7.5 s-1 at a nanoparticle concentration of 9.0 nM (0.78 mg/mL, 25 °C, 600 MHz 1H Larmor frequency). Similarly, paramagnetic rates of 10.5 s-1 were observed for water using the GdF3:cit nanoparticles at a nanoparticle concentration of 0.55 nM (0.77 mg/mL, 25 °C, 600 MHz 1H Larmor frequency). Relaxivity measurements confirmed the potential of the particles for applications as contrast agents at MRI imaging field strengths. T 1 and T 2 experiments of the GdF3:cit revealed mass relaxivities of 7.4 ± 0.2 and 8.4 ± 0.2 s-1 (mg/mL)-1, respectively, at 1.5 T, whereas identical measurements at 3.0 T revealed respective relaxivities of 8.8 ± 0.2 and 9.4 ± 0.2 s-1 (mg/mL)-1. The relatively high mass relaxivities exhibited by the nanoparticles may also find uses in NMR studies in which spin−lattice relaxation times are prohibitively long, as in the case of highly deuterated proteins. Direct interaction with the protein can be minimized by the use of surface charges opposite to the net charge of the molecule, whereas the nanoparticles are easily removed by ultracentrifugation.

This review covers current trends in studies of membrane amphiphiles and membrane proteins using both fast tumbling bicelles and magnetically aligned bicelle media for both solution state and solid state NMR. The fast tumbling bicelles provide a versatile biologically mimetic membrane model, which in many cases is preferable to micelles, both because of the range of lipids and amphiphiles that may be combined and because radius of curvature effects and strain effects common with micelles may be avoided. Drug and small molecule binding and partitioning studies should benefit from their application in fast tumbling bicelles, tailored to mimic specific membranes. A wide range of topology and immersion depth studies have been shown to be effective in fast tumbling bicelles, while residual dipolar couplings add another dimension to structure refinement possibilities, particularly for situations in which the peptide is uniformly labeled with 15N and 13C. Solid state NMR studies of polytopic transmembrane proteins demonstrate that it is possible to express, purify, and reconstitute membrane proteins, ranging in size from single transmembrane domains to seven-transmembrane GPCRs, into bicelles. The line widths and quality of the resulting 15NH dipole-15N chemical shift spectra demonstrate that there are no insurmountable obstacles to the study of large membrane proteins in magnetically aligned media.

The effects of 15 jatrophane diterpene polyesters (1-3 and 5-16) isolated from lipophilic extracts of Euphorbia serrulata, E. esula, E. salicifolia, and E. peplus (Euphorbiaceae) on the reversion of multidrug resistance of mouse lymphoma cells were examined. The structures of five new compounds (1-5) were elucidated by spectroscopic methods, including HRFABMS, ESIMS, (1)H-(1)H homonuclear and (1)H-(13)C heteronuclear correlations, long-range correlation spectra, and NOESY experiments. The stereochemistry and absolute configuration of one compound (3) were determined by X-ray crystallography. The structure-activity relationship is discussed.

The isolated N-terminal SH3 domain of the Drosophila signal transduction protein Drk (drkN SH3) is a useful model for the study of residual structure and fluctuating structure in disordered proteins since it exists in slow exchange between a folded (Fexch) and compact unfolded (Uexch) state in roughly equal proportions under nondenaturing conditions. The single tryptophan residue, Trp36, is believed to play a key role in forming a non-native hydrophobic cluster in the Uexch state, with a number of long-range nuclear Overhauser contacts (NOEs) observed primarily to the indole proton. Substitution of Trp36 for 5-fluoro-Trp36 resulted in a substantial shift in the equilibrium to favor the Fexch state. A variety of 19F NMR measurements were performed to investigate the degree of solvent exposure and hydrophobicity associated with the 5-fluoro position in both the Fexch and Uexch states. Ambient T1 measurements and H2O/D2O solvent isotope effects indicated extensive protein contacts to the 5-fluoro position in the Fexch state and greater solvent exposure in the Uexch state. This was corroborated by the measurements of paramagnetic effects (chemical shift perturbations and T1 relaxation enhancement) from dissolved oxygen at a partial pressure of 20 atm. In contrast, paramagnetic effects from dissolved oxygen revealed less solvent exposure to the indole proton of Trp36 in the Uexch state than that observed for the Fexch state, consistent with the model in which Trp36 indole belongs to a non-native cluster. Thus, although the Uexch state may be described as a dynamically interconverting ensemble of conformers, there appears to be significant asymmetry in the environment of the indole group and the six-membered ring or backbone of Trp36. This implied lack of averaging of a side chain position is in contrast to the general view of fluctuating side chains within disordered states.