Structural characterization of proteins with an attached ATCUN motif by paramagnetic relaxation enhancement NMR spectroscopy

被引：146
作者：
Donaldson, LW
Skrynnikov, NR [1 ]
Choy, WY
Muhandiram, DR
Sarkar, B
Forman-Kay, JD
Kay, LE
机构：
[1] Univ Toronto, Dept Med Genet, Toronto, ON M5S 1A8, Canada
[2] Univ Toronto, Dept Biochem, Toronto, ON M5S 1A8, Canada
[3] Univ Toronto, Dept Chem, Toronto, ON M5S 1A8, Canada
[4] Hosp Sick Children, Struct Biol & Biochem Program, Toronto, ON M5G 1X8, Canada
[5] York Univ, Dept Biol, N York, ON M3J 1P3, Canada
[6] Prot Engn Network Ctr Excellence, Kingston, ON, Canada
来源：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2001年 / 123卷 / 40期
关键词：
D O I：
10.1021/ja011241p
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
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.
引用
收藏
页码：9843 / 9847
页数：5
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