TY - JOUR
T1 - NMR structure determination and investigation using a reduced proton (REDPRO) labeling strategy for proteins
AU - Shekhtman, Alexander
AU - Ghose, Ranajeet
AU - Goger, Michael
AU - Cowburn, David
N1 - Funding Information:
We acknowledge valuable discussions with David LeMaster. We would like to thank Varian, Inc. and Oxford Instruments for allocating NMR spectrometer time and Eriks Kupce for collecting the 900 MHz data. This work was supported by NIH Grant GM-47021 and 59908 and NCI Fellowship F037244.
PY - 2002/7/31
Y1 - 2002/7/31
N2 - We present here a stable isotope labeling technique for proteins, which seeks the appropriate compromise between the advantages of (a) random isotope labeling, with its large number of protons available for structure determination, and (b) selective labeling to generate isolated proton spins decreasing spectral complexity and improving relaxation properties of NMR experiments. The described reduced proton (REDPRO) procedure results in side-chain specific protonation of overexpressed proteins, which is highly selective. The REDPRO labeling scheme provides a sufficient number of NOE constraints for structure calculation. Dramatically improved relaxation properties of the heteronuclear magnetization transfer coupled with TROSY advantages make the proposed labeling scheme an attractive approach for study of high molecular weight protein targets, their ligand sites, and interdomain interactions.
AB - We present here a stable isotope labeling technique for proteins, which seeks the appropriate compromise between the advantages of (a) random isotope labeling, with its large number of protons available for structure determination, and (b) selective labeling to generate isolated proton spins decreasing spectral complexity and improving relaxation properties of NMR experiments. The described reduced proton (REDPRO) procedure results in side-chain specific protonation of overexpressed proteins, which is highly selective. The REDPRO labeling scheme provides a sufficient number of NOE constraints for structure calculation. Dramatically improved relaxation properties of the heteronuclear magnetization transfer coupled with TROSY advantages make the proposed labeling scheme an attractive approach for study of high molecular weight protein targets, their ligand sites, and interdomain interactions.
KW - Nuclear magnetic resonance relaxation
KW - Solution structure determination
KW - Stable isotope labeling
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U2 - 10.1016/S0014-5793(02)03051-X
DO - 10.1016/S0014-5793(02)03051-X
M3 - Article
C2 - 12135763
AN - SCOPUS:0037205766
SN - 0014-5793
VL - 524
SP - 177
EP - 182
JO - FEBS Letters
JF - FEBS Letters
IS - 1-3
ER -