Abstract
Genome sequencing has shown the presence of genes coding for NO-synthase (NOS)-like proteins in bacteria. The roles and properties of these proteins remain unclear. UV-visible spectroscopy was used to characterize the recombinant NOS-like protein from Bacillus subtilis (bsNOS) in its ferric and ferrous states in the presence of various Fe(III)- and Fe(II)-heme-ligands and of a series of L-arginine (L-arg) analogs. BsNOS exhibited several spectroscopic and binding properties in common with Bacillus anthracis NOS (baNOS) that were clearly different from those of tetrahydrobiopterin (H4B)-free mammalian NOS oxygenase domains (mNOS(oxys)) and of Staphylococcus aureus NOS (saNOS). Interestingly, bsNOS and baNOS that do not contain H4B exhibited properties much closer to those of H4B-containing mNOS(oxys). Moreover, bsNOS was found to efficiently catalyze the oxidation of L-arginine into L-citrulline by H(2)O(2), whereas H4B-free mNOS(oxys) exhibited low activities for this reaction.
Copyright © 2011 Elsevier Inc. All rights reserved.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Arginine / chemistry
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Arginine / metabolism*
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Bacillus subtilis / metabolism*
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Bacteria / enzymology
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Binding Sites / genetics
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Biocatalysis
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Biopterins / analogs & derivatives
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Biopterins / metabolism
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Citrulline / chemistry
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Citrulline / metabolism
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Ferric Compounds / chemistry
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Ferric Compounds / metabolism
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Ferrous Compounds / chemistry
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Ferrous Compounds / metabolism
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Heme / chemistry
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Heme / metabolism
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Hydrogen Peroxide / chemistry
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Hydrogen Peroxide / metabolism
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Imidazoles / metabolism
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Kinetics
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Mammals / metabolism
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Nitric Oxide Synthase / chemistry
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Nitric Oxide Synthase / genetics
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Nitric Oxide Synthase / metabolism*
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Oxidation-Reduction
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Protein Binding
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Species Specificity
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Spectrophotometry
Substances
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Bacterial Proteins
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Ferric Compounds
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Ferrous Compounds
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Imidazoles
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Recombinant Proteins
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Biopterins
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Citrulline
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Heme
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Arginine
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Hydrogen Peroxide
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Nitric Oxide Synthase
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sapropterin