Structural characterization of proteins with an attached ATCUN motif by paramagnetic relaxation enhancement NMR spectroscopy
Structural characterization of proteins with an attached ATCUN motif by paramagnetic relaxation enhancement NMR spectroscopy
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DOI：
10.1021/ja011241p
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发表时间：
2001-10-10
影响因子：
15
通讯作者：
Kay, LE
中科院分区：
文献类型：
--
作者：
Donaldson, LW;Skrynnikov, NR;Kay, LE
The use of a short, three-residue Cu2+-binding sequence, the ATCUN motif, is presented as an approach for extracting long-range distance restraints from relaxation enhancement NMR spectroscopy. The ATCUN motif is prepended to the N-termini of proteins and binds Cu2+ with a very high affinity. Relaxation rates of amide protons in ATCUN-tagged protein in the presence and absence of Cu2+ can be converted into distance restraints and used for structure refinement by using a new routine, PMAG, that has been written for the structure calculation program CNS. The utility of the approach is demonstrated with an application to ATCUN-tagged ubiquitin. Excellent agreement between measured relaxation rates and those calculated on the basis of the X-ray structure of the protein have been obtained.
