Detection of a conformational change in maltose binding protein by (129)Xe NMR spectroscopy.
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ATP-Binding Cassette Transporters↗
Biomedical subjects
M M Spence
Publications and source records attributed to M M Spence.
Functionalized xenon as a biosensor.
The detection of biological molecules and their interactions is a significant component of modern biomedical research. In current biosensor technologies, simultaneous detection is limited to a small number of analytes by the spectral overlap of their signals. We have developed an NMR-based xenon biosensor that capitalizes on the enhanced signal-to-noise, spectral simplicity, and chemical-shift sensitivity of laser-polarized xenon to detect specific biomolecules at the level of tens of nanomoles. We present results using xenon "functionalized" by a biotin-modified supramolecular cage to detect biotin-avidin binding. This biosensor methodology can be extended to a multiplexing assay for multiple analytes.
Avidin↗
Characterization of the effects of nonspecific xenon-protein interactions on (129)Xe chemical shifts in aqueous solution: further development of xenon as a biomolecular probe.
The sensitivity of (129)Xe chemical shifts to weak nonspecific xenon-protein interactions has suggested the use of xenon to probe biomolecular structure and interactions. The realization of this potential necessitates a further understanding of how different macromolecular properties influence the (129)Xe chemical shift in aqueous solution. Toward this goal, we have acquired (129)Xe NMR spectra of xenon dissolved in amino acid, peptide, and protein solutions under both native and denaturing conditions. In general, these cosolutes induce (129)Xe chemical shifts that are downfield relative to the shift in water, as they deshield the xenon nucleus through weak, diffusion-mediated interactions. Correlations between the extent of deshielding and molecular properties including chemical identity, structure, and charge are reported. Xenon deshielding was found to depend linearly on protein size under denaturing solution conditions; the denaturant itself has a characteristic effect on the (129)Xe chemical shift that likely results from a change in the xenon solvation shell structure. In native protein solutions, contributions to the overall (129)Xe chemical shift arise from the presence of weak xenon binding either in cavities or at the protein surface. Potential applications of xenon as a probe of biological systems including the detection of conformational changes and the possible quantification of buried surface area at protein-protein interfaces are discussed.
Amino Acids↗
Evidence of nonspecific surface interactions between laser-polarized xenon and myoglobin in solution.
The high sensitivity of the magnetic resonance properties of xenon to its local chemical environment and the large (129)Xe NMR signals attainable through optical pumping have motivated the use of xenon as a probe of macromolecular structure and dynamics. In the present work, we report evidence for nonspecific interactions between xenon and the exterior of myoglobin in aqueous solution, in addition to a previously reported internal binding interaction. (129)Xe chemical shift measurements in denatured myoglobin solutions and under native conditions with varying xenon concentrations confirm the presence of nonspecific interactions. Titration data are modeled quantitatively with treatment of the nonspecific interactions as weak binding sites. Using laser-polarized xenon to measure (129)Xe spin-lattice relaxation times (T(1)), we observed a shorter T(1) in the presence of 1 mM denatured apomyoglobin in 6 M deuterated urea (T(1) = 59 +/- 1 s) compared with that in 6 M deuterated urea alone (T(1) = 291 +/- 2 s), suggesting that nonspecific xenon-protein interactions can enhance (129)Xe relaxation. An even shorter T(1) was measured in 1 mM apomyoglobin in D(2)O (T(1) = 15 +/- 0.3 s), compared with that in D(2)O alone (T(1) = 506 +/- 5 s). This difference in relaxation efficiency likely results from couplings between laser-polarized xenon and protons in the binding cavity of apomyoglobin that may permit the transfer of polarization between these nuclei via the nuclear Overhauser effect.
Animals↗
Training American listeners to perceive Mandarin tones.
Auditory training has been shown to be effective in the identification of non-native segmental distinctions. In this study, it was investigated whether such training is applicable to the acquisition of non-native suprasegmental contrasts, i.c., Mandarin tones. Using the high-variability paradigm, eight American learners of Mandarin were trained in eight sessions during the course of two weeks to identify the four tones in natural words produced by native Mandarin talkers. The trainees' identification accuracy revealed an average 21% increase from the pretest to the post-test, and the improvement gained in training was generalized to new stimuli (18% increase) and to new talkers and stimuli (25% increase). Moreover, the six-month retention test showed that the improvement was retained long after training by an average 21% increase from the pretest. The results are discussed in terms of non-native suprasegmental perceptual modification, and the analogies between L2 acquisition processes at the segmental and suprasegmental levels.
Adult↗
Cultural diversity process improves organizational community in urban teaching medical center.
An urban teaching facility with nearly 3,000 employees had communication problems associated with race, gender and other cultural differences. It also competed for health care dollars and faced possible reduction in federal funding. The medical center instituted mandatory training in cultural diversity and customer service-and integrated the training process with the hospital's overall quality improvement plan and marketing strategy. The integrated approach affected the bottom line-Hurley's patient base has increased, and the medical center operates in the black. Training in cultural diversity and customer service is an effective tool to improve employee communication and improve financial outlook.
Cultural Diversity↗