Structural Characterization of Proteins with an Attached ATCUN Motif by Paramagnetic Relaxation Enhancement NMR Spectroscopy
- 15 September 2001
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 123 (40), 9843-9847
- https://doi.org/10.1021/ja011241p
Abstract
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.Keywords
This publication has 27 references indexed in Scilit:
- Lanthanide-Induced Pseudocontact Shifts for Solution Structure Refinements of Macromolecules in Shells up to 40 Å from the Metal IonJournal of the American Chemical Society, 2000
- Protein global fold determination using site‐directed spin and isotope labelingProtein Science, 2000
- Binding of 15N-Labeled Isoniazid to KatG and KatG(S315T): Use of Two-Spin [zz]-Order Relaxation Rate for 15N−Fe Distance DeterminationJournal of the American Chemical Society, 1999
- Long-Range Structural Information in NMR Studies of Paramagnetic Molecules from Electron Spin−Nuclear Spin Cross-Correlated RelaxationJournal of the American Chemical Society, 1999
- The effect of axial ligand plane orientation on the contact and pseudocontact shifts of low-spin ferriheme proteinsJBIC Journal of Biological Inorganic Chemistry, 1998
- Arrays with Local Centers of Symmetry in Space Groups Pca21 and Pna21Acta crystallographica Section B, Structural science, crystal engineering and materials, 1998
- A New Method To Detect Long-Range Protein−RNA Contacts: NMR Detection of Electron−Proton Relaxation Induced by Nitroxide Spin-Labeled RNAJournal of the American Chemical Society, 1998
- NMR structure of neuromedin C, a neurotransmitter with an amino terminal CuII‐, NiII‐binding (ATCUN) motifChemical Biology & Drug Design, 1997
- Torsion-Angle Molecular Dynamics as a New Efficient Tool for NMR Structure CalculationJournal of Magnetic Resonance, 1997
- NMRPipe: A multidimensional spectral processing system based on UNIX pipesJournal of Biomolecular NMR, 1995