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.

The Development, Reliability, and Validity of the OARS Multidimensional Functional Assessment Questionnaire

This report outlines the development, validity, and reliability of Part A of the OARS Multidimensional Functional Assessment Questionnaire. Part A permits assessment of individuals' functioning on each of five dimensions (social, economic, mental health, physical health and self-care capacity), the detailed information in each area being summarized on a 6-point rating scale by a rater. Content and consensual validity were ensured by the manner of construction. Information on criterion validity was obtained for all dimensions except social. The criterion used and their associated Kendall's Tau values were: an objective economic scale (.62); ratings based on personal interviews by geropsychiatrists (.60); physician's associates (.82); and physical therapists (.89). For 11 geographically dispersed raters from research and clinic settings, intraclass correlational coefficients, based on 30 subjects, ranged from .66 on physical health to .87 in self-care capacity; 74% of the ratings were in complete agreement, 24% differed by one point.