758 resultados para Magic tricks.
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We present strategies for chemical shift assignments of large proteins by magic-angle spinning solid-state NMR, using the 21-kDa disulfide-bond-forming enzyme DsbA as prototype. Previous studies have demonstrated that complete de novo assignments are possible for proteins up to approximately 17 kDa, and partial assignments have been performed for several larger proteins. Here we show that combinations of isotopic labeling strategies, high field correlation spectroscopy, and three-dimensional (3D) and four-dimensional (4D) backbone correlation experiments yield highly confident assignments for more than 90% of backbone resonances in DsbA. Samples were prepared as nanocrystalline precipitates by a dialysis procedure, resulting in heterogeneous linewidths below 0.2 ppm. Thus, high magnetic fields, selective decoupling pulse sequences, and sparse isotopic labeling all improved spectral resolution. Assignments by amino acid type were facilitated by particular combinations of pulse sequences and isotopic labeling; for example, transferred echo double resonance experiments enhanced sensitivity for Pro and Gly residues; [2-(13)C]glycerol labeling clarified Val, Ile, and Leu assignments; in-phase anti-phase correlation spectra enabled interpretation of otherwise crowded Glx/Asx side-chain regions; and 3D NCACX experiments on [2-(13)C]glycerol samples provided unique sets of aromatic (Phe, Tyr, and Trp) correlations. Together with high-sensitivity CANCOCA 4D experiments and CANCOCX 3D experiments, unambiguous backbone walks could be performed throughout the majority of the sequence. At 189 residues, DsbA represents the largest monomeric unit for which essentially complete solid-state NMR assignments have so far been achieved. These results will facilitate studies of nanocrystalline DsbA structure and dynamics and will enable analysis of its 41-kDa covalent complex with the membrane protein DsbB, for which we demonstrate a high-resolution two-dimensional (13)C-(13)C spectrum.
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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The Attorney General’s Consumer Protection Division receives hundreds of calls and consumer complaints every year. Follow these tips to avoid unexpected expense and disappointments. This record is about: “Phishing” Scams Use New Tricks
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Performance & Development Solutions (PDS) publishes a variety of newsletters that include some great information about our programs and services. Some of the topics you may find include: Upcoming Seminars Current events or news related to training Recognition of achievements How-to section
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The MAGIC collaboration has searched for high-energy gamma-ray emission of some of the most promising pulsar candidates above an energy threshold of 50 GeV, an energy not reachable up to now by other ground-based instruments. Neither pulsed nor steady gamma-ray emission has been observed at energies of 100 GeV from the classical radio pulsars PSR J0205+6449 and PSR J2229+6114 (and their nebulae 3C58 and Boomerang, respectively) and the millisecond pulsar PSR J0218+4232. Here, we present the flux upper limits for these sources and discuss their implications in the context of current model predictions.
