Related ArticlesFirst structural investigation of the restriction ribonuclease RegB: NMR spectroscopic conditions, 13C/15N double-isotopic labelling and two-dimensional heteronuclear spectra.
Protein Expr Purif. 2004 Mar;34(1):158-65
Authors: Saďda F, Odaert B, Uzan M, Bontems F
The bacteriophage T4 genome-encoded ribonuclease RegB is the unique well-defined restriction endoribonuclease. This protein cleaves with an almost absolute specificity its RNA substrate in the middle of the GGAG tetranucleotide mainly found in the Shine-Dalgarno sequence (required for the prokaryotic initiation of the translation). This protein has no significant homology to any known ribonuclease and its structure has never been investigated. The extreme toxicity of this ribonuclease prevents the expression of large quantities for structural studies. Here, we show that the toxicity of RegB can be bypassed by using the RegB H48A point mutant and explain why resolving the structure of this mutant is relevant. For nuclear magnetic resonance (NMR) purposes, we report the preparation of highly pure (13)C/(15)N double-labelled 1.2mM samples of RegB H48A using a high yield expression procedure in minimal medium (30 mg/L). We also present a set of solution conditions that maintain the concentrated samples of this protein stable for long periods at the NMR-required temperature. Finally, we present the first (1)H/(15)N and (1)H/(13)C two-dimensional NMR spectra of RegB H48A. These spectra show that the protein is folded and that the full structural analysis of RegB by NMR is feasible.
[NMR paper] NMR studies of restriction enzyme-DNA interactions: role of conformation in sequence
NMR studies of restriction enzyme-DNA interactions: role of conformation in sequence specificity.
Related Articles NMR 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...
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[NMR paper] Structural investigation of pig metmyoglobin by 129Xe NMR spectroscopy.
Structural investigation of pig metmyoglobin by 129Xe NMR spectroscopy.
Related Articles Structural investigation of pig metmyoglobin by 129Xe NMR spectroscopy.
Biochim Biophys Acta. 2004 Sep 24;1674(2):182-92
Authors: Corda M, Era B, Fais A, Casu M
The potentiality of xenon's sensitivity to its local magnetic environment is thoroughly investigated to probe internal structural differences between pig and horse metmyoglobin (MMb). These MMb's differ by 14 amino acids. One of these, Ile142 in horse MMb, is located in the proximal cavity, which...
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11-24-2010 10:01 PM
Structural, NMR spectroscopic, and computational investigation of hemin loading in th
Structural, NMR spectroscopic, and computational investigation of hemin loading in the hemophore HasAp from Pseudomonas aeruginosa.
Related Articles Structural, NMR spectroscopic, and computational investigation of hemin loading in the hemophore HasAp from Pseudomonas aeruginosa.
J Am Chem Soc. 2010 Jul 21;132(28):9857-72
Authors: Jepkorir G, Rodríguez JC, Rui H, Im W, Lovell S, Battaile KP, Alontaga AY, Yukl ET, Moënne-Loccoz P, Rivera M
When challenged by low-iron conditions several Gram-negative pathogens secrete a hemophore (HasA) to scavenge...
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11-17-2010 05:49 PM
Towards a structural understanding of the smallest known oncoprotein: Investigation o
Towards a structural understanding of the smallest known oncoprotein: Investigation of the Bovine Papillomavirus E5 protein using solution-state NMR.
Related Articles Towards a structural understanding of the smallest known oncoprotein: Investigation of the Bovine Papillomavirus E5 protein using solution-state NMR.
Biochim Biophys Acta. 2010 Nov 9;
Authors: King G, Oates J, Patel D, van den Berg HA, Dixon AM
The homo-dimeric E5 protein from Bovine Papillomavirus activates the platelet-derived growth factor ? receptor through transmembrane (TM)...
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11-16-2010 04:13 PM
Solution structural investigation and conformation-activity relationship of BAM8-22 b
Solution structural investigation and conformation-activity relationship of BAM8-22 by NMR and molecular dynamics simulations.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Solution structural investigation and conformation-activity relationship of BAM8-22 by NMR and molecular dynamics simulations.
Bioorg Med Chem Lett. 2010 Feb 1;20(3):1260-2
Authors: Lv G, Dong S
NMR spectroscopy and molecular dynamics simulations show that BAM8-22...
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09-22-2010 05:27 AM
Restriction Endonucleases tools
Restriction Endonucleases tools
restriction endonucleases Double Digests table
Double Digest Finder
Cleavage Close to the End of DNA Fragments (oligonucleotides)
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08-25-2010 04:37 AM
[NMR paper] Three-dimensional structure of the complexes of ribonuclease A with 2',5'-CpA and 3',
Three-dimensional structure of the complexes of ribonuclease A with 2',5'-CpA and 3',5'-d(CpA) in aqueous solution, as obtained by NMR and restrained molecular dynamics.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Three-dimensional structure of the complexes of ribonuclease A with 2',5'-CpA and 3',5'-d(CpA) in aqueous solution, as obtained by NMR...
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08-22-2010 02:20 PM
[NMR paper] Investigation of ribonuclease T1 folding intermediates by hydrogen-deuterium amide ex
Investigation of ribonuclease T1 folding intermediates by hydrogen-deuterium amide exchange-two-dimensional NMR spectroscopy.
Related Articles Investigation of ribonuclease T1 folding intermediates by hydrogen-deuterium amide exchange-two-dimensional NMR spectroscopy.
Biochemistry. 1993 Jun 22;32(24):6152-6
Authors: Mullins LS, Pace CN, Raushel FM
The rate of hydrogen bond formation at individual amino acid residues in ribonuclease T1 (RNase T1) has been investigated by the hydrogen-deuterium exchange-2D NMR (HDEx-2D NMR) technique (Udgaonkar...