Source:http://linkedlifedata.com/resource/pubmed/id/12403627
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
44
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| pubmed:dateCreated |
2002-10-29
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| pubmed:abstractText |
Nitrogenase naturally requires adenosine nucleoside triphosphates and divalent metal cations for catalytic activity. Their energy of hydrolysis controls several mechanistic functions, most probably via separate structural conformers of the nitrogenase Fe protein. To characterize the ligand environment of the divalent metal in the ternary complex, with ADP or ATP and the Fe protein from Klebsiella pneumoniae, the hyperfine structures have been investigated by electron paramagnetic resonance (EPR) spectroscopy by substituting naturally occurring diamagnetic Mg(2+) by paramagnetic oxovanadium. This metal replacement leads to inhibition of nitrogenase activity. Moreover, depending on pH, two distinctly different VO(2+) EPR spectra are detected. At pH 7.4 each of the vanadyl EPR hyperfine lines is further split into two. This indicates that several spectroscopically distinguishable metal coordination environments coexist for VO(2+)-nucleotide chelate complexes in the presence of the reduced Fe protein. Overall, a total of at least three distinct local metal coordination environments have been identified. We report the EPR parameters for each of the disparate metal coordinations measured at different pH values with ADP and ATP bound. EPR spectra have also been recorded for the oxidized Fe protein showing essentially similar spectra to that of the reduced protein. The EPR parameters of VO-nucleotides in the presence of the Fe protein are consistent, for all metal coordination environments, with direct metal ligation by nucleotide phosphate groups and the formation of mononucleotide complexes. The nucleotide binding environment with the highest ligand field strength is compatible with a metal coordination structure that is also found in various G-proteins with GTP bound. No significant EPR line width change is detected after exchange into D(2)O buffer solution for any of the pH forms although differences exist between the pH forms. The missing difference between the EPR parameters in the presence of ADP or ATP suggests that there is little or no conformational rearrangement between these two forms; this contrasts with behavior of G-proteins that undergo substantial conformational changes upon hydrolysis. This could be related to the inhibition of nitrogenase by VO(2+).
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Diphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Adenosine Triphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Buffers,
http://linkedlifedata.com/resource/pubmed/chemical/Deuterium Oxide,
http://linkedlifedata.com/resource/pubmed/chemical/Enzyme Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Oxidoreductases,
http://linkedlifedata.com/resource/pubmed/chemical/Spin Labels,
http://linkedlifedata.com/resource/pubmed/chemical/Vanadates,
http://linkedlifedata.com/resource/pubmed/chemical/Vanadium Compounds,
http://linkedlifedata.com/resource/pubmed/chemical/Water,
http://linkedlifedata.com/resource/pubmed/chemical/nitrogenase reductase
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| pubmed:status |
MEDLINE
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| pubmed:month |
Nov
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| pubmed:issn |
0006-2960
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
5
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| pubmed:volume |
41
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
13253-63
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:12403627-Adenosine Diphosphate,
pubmed-meshheading:12403627-Adenosine Triphosphate,
pubmed-meshheading:12403627-Binding, Competitive,
pubmed-meshheading:12403627-Binding Sites,
pubmed-meshheading:12403627-Buffers,
pubmed-meshheading:12403627-Crystallography, X-Ray,
pubmed-meshheading:12403627-Deuterium Oxide,
pubmed-meshheading:12403627-Electron Spin Resonance Spectroscopy,
pubmed-meshheading:12403627-Enzyme Inhibitors,
pubmed-meshheading:12403627-Hydrogen-Ion Concentration,
pubmed-meshheading:12403627-Hydrolysis,
pubmed-meshheading:12403627-Klebsiella pneumoniae,
pubmed-meshheading:12403627-Magnesium,
pubmed-meshheading:12403627-Oxidoreductases,
pubmed-meshheading:12403627-Protein Conformation,
pubmed-meshheading:12403627-Spin Labels,
pubmed-meshheading:12403627-Vanadates,
pubmed-meshheading:12403627-Vanadium Compounds,
pubmed-meshheading:12403627-Water
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| pubmed:year |
2002
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| pubmed:articleTitle |
Multiple inequivalent metal-nucleotide coordination environments in the presence of the VO2+-inhibited nitrogenase iron protein: pH-dependent structural rearrangements at the nucleotide binding site.
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| pubmed:affiliation |
Department of Biological Chemistry, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK. jan.petersen@bbsrc.ac.uk
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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