Using cobalt or nickel to replace copper in native azurin allows one to fingerprint the metal coordination site of the protein. The metal sites of wild type Alcaligenes denitrificans azurin and its M121Q mutant are clearly distinguishable through the paramagnetic 1H NMR spectra of the Ni(II) and Co(II) derivatives. In the wild type azurin, Gly45 coordinates to nickel or cobalt, while Met121 appears as a weak metal ligand. On the contrary, in the M121Q azurin mutant, the metal exhibits a clear preference for the Gln121, which coordinates through the side chain carbonyl oxygen, and Gly45 is not a ligand. Changes in the isotropic shifts and relaxation properties of signals from the Cys112, His46, and His117 metal ligands suggest a movement of the metal ion out of the equatorial plane, indicating that the metal site is tetrahedral. These effects are less pronounced in the Ni(II) M121Q azurin than in the Co(II) metalloderivative. The similarity between the NMR spectra of the Co(II) derivatives of stellacyanin and the M121Q azurin is in agreement with a very similar metal site in both proteins and supports the existence of a coordinated Gln in stellacyanin.
[NMR paper] Gated electron transfers and electron pathways in azurin: a NMR dynamic study at mult
Gated electron transfers and electron pathways in azurin: a NMR dynamic study at multiple fields and temperatures.
Related Articles Gated electron transfers and electron pathways in azurin: a NMR dynamic study at multiple fields and temperatures.
J Mol Biol. 2004 Oct 1;342(5):1599-611
Authors: Zhuravleva AV, Korzhnev DM, Kupce E, Arseniev AS, Billeter M, Orekhov VY
Dynamic properties of electron transfer pathways in a small blue copper cupredoxin are explored using an extensive 15N NMR relaxation study of reduced Pseudomonas aeruginosa azurin...
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[NMR paper] Paramagnetic 1H NMR spectrum of nickel(II) pseudoazurin: investigation of the active
Paramagnetic 1H NMR spectrum of nickel(II) pseudoazurin: investigation of the active site structure and the acid and alkaline transitions.
Related Articles Paramagnetic 1H NMR spectrum of nickel(II) pseudoazurin: investigation of the active site structure and the acid and alkaline transitions.
Inorg Chem. 2002 Dec 16;41(25):6662-72
Authors: Dennison C, Sato K
The paramagnetic (1)H NMR spectrum of Ni(II) pseudoazurin possesses a number of resonances exhibiting sizable Fermi-contact shifts. These have been assigned to protons associated with...
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[NMR paper] PASE (PAramagnetic signals enhancement): a new method for NMR study of paramagnetic p
PASE (PAramagnetic signals enhancement): a new method for NMR study of paramagnetic proteins.
Related Articles PASE (PAramagnetic signals enhancement): a new method for NMR study of paramagnetic proteins.
J Magn Reson. 1998 Sep;134(1):154-7
Authors: Bondon A, Mouro C
A new method for NMR spectra acquisition of paramagnetic proteins is described, based on the simple use of homonuclear broadband decoupling of the diamagnetic region. Several advantages are associated with this method which was applied to one-dimensional spectra, to 1D...
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[NMR paper] 31P-NMR spectroscopy of human and Paracoccus denitrificans electron transfer flavopro
31P-NMR spectroscopy of human and Paracoccus denitrificans electron transfer flavoproteins, and 13C- and 15N-NMR spectroscopy of human electron transfer flavoprotein in the oxidised and reduced states.
Related Articles 31P-NMR spectroscopy of human and Paracoccus denitrificans electron transfer flavoproteins, and 13C- and 15N-NMR spectroscopy of human electron transfer flavoprotein in the oxidised and reduced states.
Eur J Biochem. 1998 Jul 1;255(1):125-32
Authors: Griffin KJ, Degala GD, Eisenreich W, Müller F, Bacher A, Frerman FE
Human and...
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[NMR paper] 1H NMR studies on the interaction of beta-carboline derivatives with human serum albu
1H NMR studies on the interaction of beta-carboline derivatives with human serum albumin.
Related Articles 1H NMR studies on the interaction of beta-carboline derivatives with human serum albumin.
J Magn Reson. 1998 Feb;130(2):281-6
Authors: Veglia G, Delfini M, Giudice MR, Gaggelli E, Valensin G
1H NMR studies were performed on two beta-carboline derivatives interacting with human serum albumin. The spin-lattice relaxation rates of the two derivatives, having side chains of different length and polarity, were used to demonstrate a diverse...
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[NMR paper] The dynamic properties of the M121H azurin metal site as studied by NMR of the parama
The dynamic properties of the M121H azurin metal site as studied by NMR of the paramagnetic Cu(II) and Co(II) metalloderivatives.
Related Articles The dynamic properties of the M121H azurin metal site as studied by NMR of the paramagnetic Cu(II) and Co(II) metalloderivatives.
J Biol Chem. 1998 Jan 2;273(1):177-85
Authors: Salgado J, Kroes SJ, Berg A, Moratal JM, Canters GW
The M121H azurin mutant in solution presents various species in equilibrium that can be detected and studied by 1H NMR of the Cu(II) and Co(II) paramagnetic...
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[NMR paper] Paramagnetic NMR Spectroscopy of Cobalt(II) and Copper(II) Derivatives of Pseudomonas
Paramagnetic NMR Spectroscopy of Cobalt(II) and Copper(II) Derivatives of Pseudomonas aeruginosa His46Asp Azurin.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles Paramagnetic NMR Spectroscopy of Cobalt(II) and Copper(II) Derivatives of Pseudomonas aeruginosa His46Asp Azurin.
Inorg Chem. 1997 Sep 24;36(20):4567-4570
Authors: Vila AJ, Ramirez BE, Di Bilio AJ, Mizoguchi TJ, Richards JH, Gray HB
NMR spectra of paramagnetic Co(II) and Cu(II) derivatives of Pseudomonas aeruginosa His46Asp...
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[NMR paper] Synthesis and interaction studies of 13C labeled lactone derivatives with a model pro
Synthesis and interaction studies of 13C labeled lactone derivatives with a model protein using 13C NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Synthesis and interaction studies of 13C labeled lactone derivatives with a model protein using 13C NMR.
Bioorg Med Chem. 1993 Nov;1(5):389-97
Authors: Franot C, Benezra C, Lepoittevin JP
Two molecules 9 and 14, representatives of two series of electrophilic lactone derivatives, have been synthesized, and labeled with...