Related Articles1H NMR studies on the CuA center of nitrous oxide reductase from Pseudomonas stutzeri.
Biochemistry. 1999 Aug 24;38(34):11164-71
Authors: Holz RC, Alvarez ML, Zumft WG, Dooley DM
1H NMR spectra of the CuA center of N2OR from Pseudomonas stutzeri, and a mutant enzyme that contains only CuA, were recorded in both H2O- and D2O-buffered solution at pH 7.5. Several sharp, well-resolved hyperfine-shifted 1H NMR signals were observed in the 60 to -10 ppm chemical shift range. Comparison of the native and mutant N2OR spectra recorded in H2O-buffered solutions indicated that several additional signals are present in the native protein spectrum. These signals are attributed to a dinuclear copperII center. At least two of the observed hyperfine-shifted signals associated with the dinuclear center, those at 23.0 and 13.2 ppm, are lost upon replacement of H2O buffer with D2O buffer. These data indicate that at least two histidine residues are ligands of a dinuclear CuII center. Comparison of the mutant N2OR 1H NMR spectra recorded in H2O and D2O indicates that three signals, c (27.5 ppm), e (23.6 ppm), and i (12.4 ppm), are solvent exchangeable. The two most strongly downfield-shifted signals (c and e) are assigned to the two N epsilon 2H (N-H) protons of the coordinated histidine residues, while the remaining exchangeable signal is assigned to a backbone N-H proton in close proximity to the CuA cluster. Signal e was found to decrease in intensity as the temperature was increased, indicating that proton e resides on a more solvent-exposed histidine residue. One-dimensional nOe studies at pH 7.5 allowed the histidine ring protons to be definitively assigned, while the remaining signals were assigned by comparison to previously reported spectra from CuA centers. The temperature dependence of the observed hyperfine-shifted 1H NMR signals of mutant N2OR were recorded over the temperature range of 276-315 K. Both Curie and anti-Curie temperature dependencies are observed for sets of hyperfine-shifted protons. Signals a and h (cysteine protons) follow anti-Curie behavior (contact shift increases with increasing temperatures), while signals b-g, i, and j (histidine protons) follow Curie behavior (contact shift decreases with increasing temperatures). Fits of the temperature dependence of the observed hyperfine-shifted signals provided the energy separation (Delta EL) between the ground (2B3u) and excited (2B2u) states. The temperature data obtained for all of the observed hyperfine-shifted histidine ligand protons provided a Delta EL value of 62 +/- 35 cm-1. The temperature dependence of the observed cysteine C beta H and C alpha H protons (a and h) were fit in a separate experiment providing a Delta EL value of 585 +/- 125 cm-1. The differences between the Delta EL values determined by 1H NMR spectroscopy and those determined by EPR or MCD likely arise from coupling between relatively low-frequency vibrational states and the ground and excited electronic states.
[NMR paper] Kinetic, Raman, NMR, and site-directed mutagenesis studies of the Pseudomonas sp. str
Kinetic, Raman, NMR, and site-directed mutagenesis studies of the Pseudomonas sp. strain CBS3 4-hydroxybenzoyl-CoA thioesterase active site.
Related Articles Kinetic, Raman, NMR, and site-directed mutagenesis studies of the Pseudomonas sp. strain CBS3 4-hydroxybenzoyl-CoA thioesterase active site.
Biochemistry. 2002 Sep 17;41(37):11152-60
Authors: Zhuang Z, Song F, Zhang W, Taylor K, Archambault A, Dunaway-Mariano D, Dong J, Carey PR
4-Hydroxybenzoyl-coenzyme A (4-HBA-CoA) thioesterase catalyzes the hydrolysis of 4-HBA-CoA to 4-hydroxybenzoate...
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[NMR paper] Solution conformation of cytochrome c-551 from Pseudomonas stutzeri ZoBell determined
Solution conformation of cytochrome c-551 from Pseudomonas stutzeri ZoBell determined by NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-cellhub.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Solution conformation of cytochrome c-551 from Pseudomonas stutzeri ZoBell determined by NMR.
Biophys J. 1994 Sep;67(3):1207-15
Authors: Cai M, Timkovich R
1H NMR spectroscopy and solution structure computations have...
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[NMR paper] Studies on ribonucleoside-diphosphate reductase from Escherichia coli. The product dC
Studies on ribonucleoside-diphosphate reductase from Escherichia coli. The product dCDP is a competitive inhibitor and functions as a spectroscopic probe for the substrate binding site; demonstration by enzyme kinetics and 1H NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles Studies on ribonucleoside-diphosphate reductase from Escherichia coli. The product dCDP is a competitive inhibitor and functions as a spectroscopic probe for the substrate binding site;...
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[NMR paper] NMR studies of Azotobacter vinelandii and Pseudomonas putida seven-iron ferredoxins.
NMR studies of Azotobacter vinelandii and Pseudomonas putida seven-iron ferredoxins. Direct assignment of beta-cysteinyl carbon NMR resonances and further proton NMR assignments of cysteinyl and aromatic resonances.
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J Biol Chem. 1992 Apr 25;267(12):8073-80
Authors: Cheng H, Grohmann K, Sweeney W
Ferredoxins are...
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[NMR paper] In vivo 13C and 15N NMR studies of methylamine metabolism in Pseudomonas species MA.
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J Biol Chem. 1991 Jun 25;266(18):11705-13
Authors: Jones JG, Bellion E
Pseudomonas species MA was grown with methylamine as a sole source of carbon and nitrogen enabling the total flow of carbon and nitrogen into this organism to be simultaneously monitored in vivo using 13C and 15N NMR. Methylamine was rapidly and extensively incorporated into the methyl...
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[NMR paper] NMR studies on p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens and salicyl
NMR studies on p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens and salicylate hydroxylase from Pseudomonas putida.
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Eur J Biochem. 1991 Sep 15;200(3):731-8
Authors: Vervoort J, Van Berkel WJ, Müller F, Moonen CT
p-Hydroxybenzoate hydroxylase from...
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[NMR paper] NMR studies on p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens and salicyl
NMR studies on p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens and salicylate hydroxylase from Pseudomonas putida.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles NMR studies on p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens and salicylate hydroxylase from Pseudomonas putida.
Eur J Biochem. 1991 Sep 15;200(3):731-8
Authors: Vervoort J, Van Berkel WJ, Müller F, Moonen CT
p-Hydroxybenzoate hydroxylase from...
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[NMR paper] NMR studies of interactions of ligands with dihydrofolate reductase.
NMR studies of interactions of ligands with dihydrofolate reductase.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles NMR studies of interactions of ligands with dihydrofolate reductase.
Biochem Pharmacol. 1990 Jul 1;40(1):141-52
Authors: Feeney J
NMR spectroscopy is a useful technique for studying interactions, conformations and dynamic processes within ligand-protein complexes. Several examples of the application of the method to studies of complexes of anti-folate...