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Proton evolved local field solid-state nuclear magnetic resonance using Hadamard encoding: theory and application to membrane proteins.

ABSTRACT: NMR anisotropic parameters such as dipolar couplings and chemical shifts are central to structure and orientation determination of aligned membrane proteins and liquid crystals. Among the separated local field experiments, the proton evolved local field (PELF) scheme is particularly suitable to measure dynamically averaged dipolar couplings and give information on local molecular motions. However, the PELF experiment requires the acquisition of several 2D datasets at different mixing times to optimize the sensitivity for the complete range of dipolar couplings of the resonances in the spectrum. Here, we propose a new PELF experiment that takes the advantage of the Hadamard encoding (HE) to obtain higher sensitivity for a broad range of dipolar couplings using a single 2D experiment. The HE scheme is obtained by selecting the spin operators with phase switching of hard pulses. This approach enables one to detect four spin operators, simultaneously, which can be processed into two 2D spectra covering a broader range of dipolar couplings. The advantages of the new approach are illustrated for a U-(15)N NAL single crystal and the U-(15)N labeled single-pass membrane protein sarcolipin reconstituted in oriented lipid bicelles. The HE-PELF scheme can be implemented in other multidimensional experiments to speed up the characterization of the structure and dynamics of oriented membrane proteins and liquid crystalline samples.

SUBMITTER: Gopinath T

PROVIDER: S-EPMC3172034 | biostudies-other | 2011 Aug

REPOSITORIES: biostudies-other

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Project description:PurposeThe purpose of this work was to examine the suitability of VIPARnd polymer gel-9.4 T magnetic resonance microimaging system for high spatial resolution dose distribution measurements.MethodsThe VIPARnd samples (3 cm in outside diameter and 12 cm in height) were exposed to ionizing radiation by using a linear accelerator (Varian TrueBeam, USA; 6 MV x-ray beam). In the calibration stage, nine gel dosimeter vials were irradiated in a water phantom homogenously to the doses from 1.5 to 30 Gy in order to obtain R2-dose relation. In the verification stage, two gel dosimeter vials were irradiated in the half beam penumbra area of 10 × 10 cm radiation field using the amount of monitor units appropriate to deliver 20 Gy at the field center. The gels were imaged on a vertical 9.4 T magnetic resonance (MR) microimaging scanner using single slice and multislice (9 slices) multiecho (90 × 7 ms) sequences at the spatial resolutions of 0.2-0.4 × 0.2-0.4 × 3 mm3 and 0.2-0.4 × 0.2-0.4 × 1 mm3 respectively. The gels were subjected to microimaging during the period of two weeks after irradiation. The reference data consisted of the dose profiles measured using the diode dosimetry, radiochromic film, ionization chamber, and the water phantom system.ResultsThe VIPARnd -9.4 T MR microimaging system was characterized by the dose sensitivity of 0.067 ± 0.002 Gy-1 s-1 at day 3 after irradiation. The dose resolution at 10 Gy (at P = 95%) was equal to 0.42 Gy at day 3 after irradiation using a single slice sequence (0.2 × 0.2 × 3 mm3 ) and 2.0 Gy at day 4 after irradiation using a multislice sequence (0.2 × 0.2 × 1 mm3 ) for one signal acquisition (measurement time: 15 min). These values were improved by ~1.4-fold when using four signal acquisitions in the single slice sequence, and by ~2.78-fold for 12 signal acquisitions in the multislice sequence. Furthermore, decreasing the in-plane resolution from 0.2 × 0.2 mm2 to 0.4 × 0.4 mm2 resulted in a dose resolution of 0.3 Gy and 1 Gy at 10 Gy (at P = 95%) for one signal acquisition in the single slice and multislice sequences respectively (measurement time: 7.5 min). As reveals from the gamma index analysis the dose distributions measured at days 3-4 postirradiation using both VIPARnd verification phantoms agree with the data obtained using a silicon diode, assuming 1 mm/5% criterion. A good interphantom reproducibility of the polymer gel dosimetry was proved by monitoring of two phantoms up to 10 days after irradiation. However, the agreement between the dose distributions measured using the diode and polymer gel started to get worse from day 5 after irradiation.ConclusionThe VIPARnd -9.4T MR microimaging system allows to obtain dose resolution of 0.42 Gy at 10 Gy (at P = 95%) for a spatial resolution of 0.2 × 0.2 × 3 mm3 (acquisition time: 15 min). Further studies are required to improve a temporal stability of the gel-derived dose distribution.

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Proton evolved local field solid-state nuclear magnetic resonance using Hadamard encoding: theory and application to membrane proteins.

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