Related ArticlesNMR studies of restriction enzyme-DNA interactions: role of conformation in sequence specificity.
Biochemistry. 2005 Apr 5;44(13):5065-74
Authors: Dupureur CM
Sequence specific DNA binding proteins are thought to adopt distinct conformations when binding to target (cognate) and nontarget (noncognate) sequences. There is both biochemical and crystallographic evidence that this behavior is important in mediating sequence recognition by the Mg(II)-dependent type II restriction enzymes. Despite this, there are few systematic comparisons of the structural behavior of these enzymes in various complexes. Here, (1)H-(15)N HSQC NMR spectroscopy is applied to PvuII endonuclease (2 x 18 kDa) in an effort to better understand the relationship between sequence recognition and enzyme conformational behavior. Spectra of the free enzyme collected in the absence and presence of metal ions indicate that while there is a modest backbone conformational response upon binding Ca(II), this does not occur with Mg(II). Substrate binding itself is accompanied by very dramatic spectral changes consistent with a large-scale conformational response. HSQC spectra of the enzyme bound to cognate (specific) and noncognate (nonspecific) oligonucleotides in the presence of Ca(II) are dramatically distinct, revealing for the first time the structural uniqueness of a PvuII cognate complex in solution. The strong correlation between NMR spectral overlap and crystallographic data (C(alpha) rmsd) permits characterization of the nonspecific PvuII complex as being more similar to the free enzyme than to the specific complex. Collectively, these data support the notion that it is the DNA, not the metal ion, which promotes a unique conformational response by the enzyme. It therefore follows that the principle role of metal ions in complex formation is one of driving substrate affinity and stability rather than conformationally priming the enzyme for substrate binding and sequence recognition. These results not only provide valuable insights into the mechanism of protein-DNA interactions but also demonstrate the utility of NMR spectroscopy in structure-function studies of these representative nucleic acid systems.
Modular protein-RNA interactions regulating mRNA metabolism: a role for NMR.
Modular protein-RNA interactions regulating mRNA metabolism: a role for NMR.
Modular protein-RNA interactions regulating mRNA metabolism: a role for NMR.
Eur Biophys J. 2011 Apr 7;
Authors: Cukier CD, Ramos A
Here we review the role played by transient interactions between multi-functional proteins and their RNA targets in the regulation of mRNA metabolism, and we describe the important function of NMR spectroscopy in the study of these systems. We place emphasis on a general approach for the study of different features of modular multi-domain...
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04-08-2011 10:00 AM
[NMR paper] CD and NMR studies of prion protein (PrP) helix 1. Novel implications for its role in
CD and NMR studies of prion protein (PrP) helix 1. Novel implications for its role in the PrPC-->PrPSc conversion process.
Related Articles CD and NMR studies of prion protein (PrP) helix 1. Novel implications for its role in the PrPC-->PrPSc conversion process.
J Biol Chem. 2003 Dec 12;278(50):50175-81
Authors: Ziegler J, Sticht H, Marx UC, Müller W, Rösch P, Schwarzinger S
The conversion of prion helix 1 from an alpha-helical into an extended conformation is generally assumed to be an essential step in the conversion of the cellular isoform...
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11-24-2010 09:16 PM
[NMR paper] DNA bending and sequence-dependent backbone conformation NMR and computer experiments
DNA bending and sequence-dependent backbone conformation NMR and computer experiments.
Related Articles DNA bending and sequence-dependent backbone conformation NMR and computer experiments.
Eur J Biochem. 1999 Oct 1;265(1):35-53
Authors: Ojha RP, Dhingra MM, Sarma MH, Shibata M, Farrar M, Turner CJ, Sarma RH
Although DNA bending plays a crucial role in several biological processes, very little is known experimentally about the relationship between sugar phosphate conformation and sequence directed bending. In this paper, we determine the...
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11-18-2010 08:31 PM
[NMR paper] A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein r
A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
FEBS Lett. 1997 Jun 9;409(2):227-31
Authors: Tsuda S, Ito A, Matsushima N
The 1H-NMR spectrum of a synthetic 24-residue peptide (A1-G-V-D-S-S-L-I-A-G-Y-G-S-T-Q-T-S-G-S-D-S-A-L-T24; INP24),...
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08-22-2010 03:31 PM
[NMR paper] A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein r
A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy.
FEBS Lett. 1997 Jun 9;409(2):227-31
Authors: Tsuda S, Ito A, Matsushima N
The 1H-NMR spectrum of a synthetic 24-residue peptide (A1-G-V-D-S-S-L-I-A-G-Y-G-S-T-Q-T-S-G-S-D-S-A-L-T24; INP24),...
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08-22-2010 03:03 PM
[NMR paper] NMR studies of retinoid-protein interactions: the conformation of [13C]-beta-ionones
NMR studies of retinoid-protein interactions: the conformation of -beta-ionones bound to beta-lactoglobulin B.
Related Articles NMR studies of retinoid-protein interactions: the conformation of -beta-ionones bound to beta-lactoglobulin B.
Pharm Res. 1999 May;16(5):651-9
Authors: Curley RW, Sundaram AK, Fowble JW, Abildgaard F, Westler WM, Markley JL
PURPOSE: Vitamin A (retinol) and its metabolites comprise the natural retinoids. While the biological action of these molecules are thought to be primarily mediated by ca. 55 kDa nuclear retinoic...
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08-21-2010 04:03 PM
Prediction of Xaa-Pro peptide bond conformation from sequence and chemical shifts
Abstract We present a program, named Promega, to predict the Xaa-Pro peptide bond conformation on the basis of backbone chemical shifts and the amino acid sequence. Using a chemical shift database of proteins of known structure together with the PDB-extracted amino acid preference of cis Xaa-Pro peptide bonds, a cis/trans probability score is calculated from the backbone and 13Cβ chemical shifts of the proline and its neighboring residues. For an arbitrary number of input chemical shifts, which may include Pro-13Cγ, Promega calculates the statistical probability that a Xaa-Pro peptide bond...
NMR studies of restriction enzyme-DNA interactions: role of conformation in sequence
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