Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Oct 11;49(42):7803-6.
doi: 10.1002/anie.201002044.

Dynamic nuclear polarization of deuterated proteins

Ümit Akbey  1 , W Trent Franks, Arne Linden, Sascha Lange, Robert G Griffin, Barth-Jan van Rossum, Hartmut Oschkinat
Affiliations

Dynamic nuclear polarization of deuterated proteins

Ümit Akbey et al. Angew Chem Int Ed Engl. .
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
The pulse sequences used to record the DNP enhanced 13C spectra with A) CP-MAS and B) direct 13C excitation, at approximately 98 K and approximately 9 kHz MAS. C) 13C CP-MAS spectrum of protonated SH3 with DNP. DNP enhanced D) 13C CP-MAS and E),F) MAS spectra of the deuterated-SH3 with 50% exchangeable proton content. A relaxation delay (RD) of 2 s (C–E) and 12 s (F) were used. Continuous-wave microwave irradiation was used while acquiring the DNP enhanced spectra. For calculation of the DNP enhancement, the spectra were recorded with microwave irradiation and compared to the spectra recorded without microwave irradiation under exactly same experimental conditions. The spectra are plotted with the same noise level, to allow direct comparison.
Figure 2
Figure 2
The dependence of the A) DNP enhancement and B) T1 relaxation time (seconds) on the overall 1H content. For comparison, a fully protonated (at exchangeable and non-exchangeable sites) and three different deuterated SH3 (at non-exchangeable sites) proteins were used. The proton content at the exchangeable sites of the protein was tuned by recrystallization of SH3 in buffers containing different H2O/D2O ratios. The H2O contents were set around 15, 25, 50 and 100% to be able to cover the full range of exchangeable proton content. The spectra were recorded at a temperature of approximately 98 K, an MAS frequency of approximately 9 kHz, and with approximately 5 W microwave irradiation in zirconium rotors. The experimental data points are connected with lines as a guide to the eye, the data points are discontinuous.
Figure 3
Figure 3
The dependence of the 1H, 13C, and 15N DNP enhancements on temperature. Results for the protonated and perdeuterated (15 and 50% exchangeable proton content) SH3 are shown. Enhancement values are calculated at each temperature for each type of experiment. For the protonated SH3, only the 13C CPMAS DNP enhancement values are shown.
See this image and copyright information in PMC

References

    1. Griffin RG. Nat. Struct. Biol. 1998;5:508. - PubMed
    1. McDermott AE. Curr. Opin. Struct. Biol. 2004;14:554. - PubMed
    1. Tycko R. Quart. Rev. Biophys. 2006;39:1. - PubMed
    1. Jaroniec CP, MacPhee CE, Bajaj VS, McMahon MT, Dobson CM, Griffin RG. Proc. Natl. Acad. Sci. USA. 2004;101:711. - PMC - PubMed
    1. Wasmer C, Lange A, Van Melckebeke H, Siemer AB, Riek R, Meier BH. Science. 2008;319:1523. - PubMed

LinkOut - more resources