TY - JOUR
T1 - Accelerated acquisition of high resolution triple-resonance spectra using non-uniform sampling and maximum entropy reconstruction
AU - Rovnyak, David
AU - Frueh, Dominique P.
AU - Sastry, Mallika
AU - Sun, Zhen Yu J.
AU - Stern, Alan S.
AU - Hoch, Jeffrey C.
AU - Wagner, Gerhard
N1 - Funding Information:
We acknowledge support from the National Institute of Health (Grants GM47467 and RR 00995 to G.W., CA89940 to D.R.) and the Rowland Institute at Harvard. We thank Drs. Thilo Stehle and Gowtham Subbarao for providing the protein sample. We thank Dr. Sven Hyberts for help with the program IBIS and Gregory Heffron for assistance with the NMR instrument. DF was partially supported by the Swiss National Science Foundation (81EL-068377).
PY - 2004/9
Y1 - 2004/9
N2 - Non-uniform sampling is shown to provide significant time savings in the acquisition of a suite of three-dimensional NMR experiments utilized for obtaining backbone assignments of H, N, C′, CA, and CB nuclei in proteins: HNCO, HN(CA)CO, HNCA, HN(CO)CA, HNCACB, and HN(CO)CACB. Non-uniform sampling means that data were collected for only a subset of all incremented evolution periods, according to a user-specified sampling schedule. When the suite of six 3D experiments was acquired in a uniform fashion for an 11kDa cytoplasmic domain of a membrane protein at 1.5mM concentration, a total of 146h was consumed. With non-uniform sampling, the same experiments were acquired in 32h and, through subsequent maximum entropy reconstruction, yielded spectra of similar quality to those obtained by conventional Fourier transform of the uniformly acquired data. The experimental time saved with this methodology can significantly accelerate protein structure determination by NMR, particularly when combined with the use of automated assignment software, and enable the study of samples with poor stability at room temperature. Since it is also possible to use the time savings to acquire a greater numbers of scans to increase sensitivity while maintaining high resolution, this methodology will help extend the size limit of proteins accessible to NMR studies, and open the way to studies of samples that suffer from solubility problems.
AB - Non-uniform sampling is shown to provide significant time savings in the acquisition of a suite of three-dimensional NMR experiments utilized for obtaining backbone assignments of H, N, C′, CA, and CB nuclei in proteins: HNCO, HN(CA)CO, HNCA, HN(CO)CA, HNCACB, and HN(CO)CACB. Non-uniform sampling means that data were collected for only a subset of all incremented evolution periods, according to a user-specified sampling schedule. When the suite of six 3D experiments was acquired in a uniform fashion for an 11kDa cytoplasmic domain of a membrane protein at 1.5mM concentration, a total of 146h was consumed. With non-uniform sampling, the same experiments were acquired in 32h and, through subsequent maximum entropy reconstruction, yielded spectra of similar quality to those obtained by conventional Fourier transform of the uniformly acquired data. The experimental time saved with this methodology can significantly accelerate protein structure determination by NMR, particularly when combined with the use of automated assignment software, and enable the study of samples with poor stability at room temperature. Since it is also possible to use the time savings to acquire a greater numbers of scans to increase sensitivity while maintaining high resolution, this methodology will help extend the size limit of proteins accessible to NMR studies, and open the way to studies of samples that suffer from solubility problems.
KW - Maximum entropy reconstruction
KW - Non-uniform sampling
KW - Protein backbone chemical shift assignments
KW - Reduced time multidimensional NMR spectroscopy
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U2 - 10.1016/j.jmr.2004.05.016
DO - 10.1016/j.jmr.2004.05.016
M3 - Article
C2 - 15324754
AN - SCOPUS:4243138246
SN - 1090-7807
VL - 170
SP - 15
EP - 21
JO - Journal of Magnetic Resonance
JF - Journal of Magnetic Resonance
IS - 1
ER -