Related ArticlesBinding ability of a HHP-tagged protein towards Ni2+ studied by paramagnetic NMR relaxation: the possibility of obtaining long-range structure information.
The binding ability of a protein with a metal binding tag towards Ni(2+) was investigated by longitudinal paramagnetic NMR relaxation, and the possibility of obtaining long-range structure information from the paramagnetic relaxation was explored. A protein with a well-defined solution structure (Escherichia coli thioredoxin) was used as the model system, and the peptide His-His-Pro (HHP) fused to the N-terminus of the protein was used as the metal binding tag. It was found that the tag forms a stable dimer complex with the paramagnetic Ni(2+) ion, where each metal ion binds two HHP-tagged protein molecules. However, it was also found that additional sites in the protein compete with the HHP-tag for the binding of the metal ion. These binding sites were identified as the side chain carboxylate groups of the aspartic and glutamic acid residues. Yet, the carboxylate groups bind the Ni(2+) ions considerably weaker than the HHP-tag, and only protons spatially close to the carboxylate sites are affected by the Ni(2+) ions bound to these groups. As for the protons that are unaffected by the carboxylate-bound Ni(2+) ions, it was found that the long-range distances derived from the paramagnetic relaxation enhancements are in good agreement with the solution structure of thioredoxin. Specifically, the obtained long-range paramagnetic distance constraints revealed that the dimer complex is asymmetric with different orientations of the two protein molecules relative to the Ni(2+) ion.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Chem Commun (Camb). 2011 May 27;
Authors: Swarbrick JD, Ung P, Su XC, Maleckis A, Chhabra S, Huber T, Otting G, Graham B
Attachment of two nitrilotriacetic acid-based ligands to a protein ?-helix in an i, i + 4 configuration produces an octadentate chelating motif that is able to bind paramagnetic...
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05-28-2011 06:50 PM
[NMR paper] Metal binding sites in proteins: identification and characterization by paramagnetic NMR relaxation.
Metal binding sites in proteins: identification and characterization by paramagnetic NMR relaxation.
Related Articles Metal binding sites in proteins: identification and characterization by paramagnetic NMR relaxation.
Biochemistry. 2005 Aug 23;44(33):11014-23
Authors: Jensen MR, Petersen G, Lauritzen C, Pedersen J, Led JJ
A method is presented that allows the identification and quantitative characterization of metal binding sites in proteins using paramagnetic nuclear magnetic resonance spectroscopy. The method relies on the nonselective...
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12-01-2010 06:56 PM
[NMR paper] The energetic cost of domain reorientation in maltose-binding protein as studied by N
The energetic cost of domain reorientation in maltose-binding protein as studied by NMR and fluorescence spectroscopy.
Related Articles The energetic cost of domain reorientation in maltose-binding protein as studied by NMR and fluorescence spectroscopy.
Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12700-5
Authors: Millet O, Hudson RP, Kay LE
Maltose-binding protein (MBP) is a two-domain protein that undergoes a ligand-mediated conformational rearrangement from an "open" to a "closed" structure on binding to maltooligosaccharides. To...
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11-24-2010 09:16 PM
[NMR paper] Protein dynamics using frequency-dependent order parameters from analysis of NMR rela
Protein dynamics using frequency-dependent order parameters from analysis of NMR relaxation data.
Related Articles Protein dynamics using frequency-dependent order parameters from analysis of NMR relaxation data.
J Magn Reson. 2003 Mar;161(1):118-25
Authors: Idiyatullin D, Daragan VA, Mayo KH
A novel approach is described to analyze NMR relaxation data on proteins. This method introduces the frequency-dependent order parameter, S(2)(omega), in order to estimate contributions to the generalized order parameter S(2) from different motional...
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11-24-2010 09:01 PM
[NMR paper] Ligand-induced structural changes to maltodextrin-binding protein as studied by solut
Ligand-induced structural changes to maltodextrin-binding protein as studied by solution NMR spectroscopy.
Related Articles Ligand-induced structural changes to maltodextrin-binding protein as studied by solution NMR spectroscopy.
J Mol Biol. 2001 Jun 15;309(4):961-74
Authors: Evenäs J, Tugarinov V, Skrynnikov NR, Goto NK, Muhandiram R, Kay LE
Solution NMR studies on the physiologically relevant ligand-free and maltotriose-bound states of maltodextrin-binding protein (MBP) are presented. Together with existing data on MBP in complex with...
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11-19-2010 08:32 PM
[NMR paper] An investigation of the dynamics of ribosomal protein L9 using heteronuclear NMR rela
An investigation of the dynamics of ribosomal protein L9 using heteronuclear NMR relaxation measurements.
Related Articles An investigation of the dynamics of ribosomal protein L9 using heteronuclear NMR relaxation measurements.
J Mol Biol. 1998 Aug 21;281(3):539-51
Authors: Lillemoen J, Hoffman DW
The dynamic properties of ribosomal protein L9 from Bacillus stearothermophilus were investigated in solution using an analysis of nitrogen-15 longitudinal and transverse relaxation rates and amide nitrogen-proton nuclear Overhauser effects. The...
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[NMR paper] Effect of antibody binding on protein motions studied by hydrogen-exchange labeling a
Effect of antibody binding on protein motions studied by hydrogen-exchange labeling and two-dimensional NMR.
Related Articles Effect of antibody binding on protein motions studied by hydrogen-exchange labeling and two-dimensional NMR.
Biochemistry. 1992 Nov 10;31(44):10678-85
Authors: Mayne L, Paterson Y, Cerasoli D, Englander SW
We have used hydrogen-exchange labeling detected by 2D NMR to study antibody-protein interactions for two monoclonal antibodies raised against horse cytochrome c. The data show that these antibodies bind mainly to the...
Binding ability of a HHP-tagged protein towards Ni2+ studied by paramagnetic NMR rela
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