Enhanced Sensitivity by Nonuniform Sampling Enables Multidimensional MAS NMR Spectroscopy of Protein Assemblies

We report dramatic sensitivity enhancements in multidimensional MAS NMR spectra by the use of nonuniform sampling (NUS) and introduce maximum entropy interpolation (MINT) processing that assures the linearity between the time and frequency domains of the NUS acquired data sets. A systematic analysis of sensitivity and resolution in 2D and 3D NUS spectra reveals that with NUS, at least 1.5- to 2-fold sensitivity enhancement can be attained in each indirect dimension without compromising the spectral resolution. These enhancements are similar to or higher than those attained by the newest-generation commercial cryogenic probes. We explore the benefits of this NUS/MaxEnt approach in proteins and protein assemblies using 1-73-(U-C-13,N-15)/74-108-(U-N-15) Escherichia coil thioredoxin reassembly. We demonstrate that in thioredoxin reassembly, NUS permits acquisition of high-quality 3D-NCACX spectra, which are inaccessible with conventional sampling due to prohibitively long experiment times. Of critical importance, issues that hinder NUS-based SNR enhancement in 3D-NMR of liquids are mitigated in the study of solid samples in which theoretical enhancements on the order of 3-4 fold are accessible by compounding the NUS-based SNR enhancement of each indirect dimension. NUS/MINT is anticipated to be widely applicable and advantageous for multidimensional heteronuclear MAS NMR spectroscopy of proteins, protein assemblies, and other biological systems.

Chemical Compatibility of Model Soil-Bentonite Backfill Containing Multiswellable Bentonite

The objective of this study was to evaluate the chemical compatibility of model soil-bentonite backfills containing multiswellable bentonite (MSB) relative to that of similar backfills containing untreated sodium (Na) bentonite or a commercially available, contaminant resistant bentonite (SW101). Flexible-wall tests were conducted on consolidated backfill specimens (effective stress =34.5 kPa) containing clean sand and 4.5–5.7% bentonite (by dry weight) using tap water and calcium chloride (CaCl2) solutions (10–1,000 mM) as the permeant liquids. Final values of hydraulic conductivity (k) and intrinsic permeability (K) to the CaCl2 solutions were determined after achieving both short-term termination criteria as defined by ASTM D5084 and long-term termination criteria for chemical equilibrium between the influent and effluent. Specimens containing MSB exhibited the smallest increases in k and K upon permeation with a given CaCl2 solution relative to specimens containing untreated Na bentonite or SW101. However, none of the specimens exhibited more than a five-fold increase in k or K, regardless of CaCl2 concentration or bentonite type. Final k values for specimens permeated with a given CaCl2 solution after permeation with tap water were similar to those for specimens of the same backfill permeated with only the CaCl2 solution, indicating that the order of permeation had no significant effect on k. Also, final k values for all specimens were within a factor of two of the k measured after achieving the ASTM D5084 termination criteria. Thus, use of only the ASTM D5084 criteria would have been sufficient to obtain reasonable estimates of long-term hydraulic conductivity for the specimens in this study.