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Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins

Physarum polycephalum expresses two closely related, calcium-independent NOSs (nitric oxide synthases). In our previous work, we showed that both NOSs are induced during starvation and apparently play a functional role in sporulation. In the present study, we characterized the genomic structures of...

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Autores principales: Messner, Simon, Leitner, Stephan, Bommassar, Christian, Golderer, Georg, Gröbner, Peter, Werner, Ernst R., Werner-Felmayer, Gabriele
Formato: Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2677215/
https://www.ncbi.nlm.nih.gov/pubmed/19046139
http://dx.doi.org/10.1042/BJ20080192
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author Messner, Simon
Leitner, Stephan
Bommassar, Christian
Golderer, Georg
Gröbner, Peter
Werner, Ernst R.
Werner-Felmayer, Gabriele
author_facet Messner, Simon
Leitner, Stephan
Bommassar, Christian
Golderer, Georg
Gröbner, Peter
Werner, Ernst R.
Werner-Felmayer, Gabriele
author_sort Messner, Simon
collection PubMed
description Physarum polycephalum expresses two closely related, calcium-independent NOSs (nitric oxide synthases). In our previous work, we showed that both NOSs are induced during starvation and apparently play a functional role in sporulation. In the present study, we characterized the genomic structures of both Physarum NOSs, expressed both enzymes recombinantly in bacteria and characterized their biochemical properties. Whereas the overall genomic organization of Physarum NOS genes is comparable with various animal NOSs, none of the exon–intron boundaries are conserved. Recombinant expression of clones with various N-termini identified N-terminal amino acids essential for enzyme activity, but not required for haem binding or dimerization, and suggests the usage of non-AUG start codons for Physarum NOSs. Biochemical characterization of the two Physarum isoenzymes revealed different affinities for L-arginine, FMN and 6R-5,6,7,8-tetrahydro-L-biopterin.
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spelling pubmed-26772152009-05-07 Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins Messner, Simon Leitner, Stephan Bommassar, Christian Golderer, Georg Gröbner, Peter Werner, Ernst R. Werner-Felmayer, Gabriele Biochem J Research Article Physarum polycephalum expresses two closely related, calcium-independent NOSs (nitric oxide synthases). In our previous work, we showed that both NOSs are induced during starvation and apparently play a functional role in sporulation. In the present study, we characterized the genomic structures of both Physarum NOSs, expressed both enzymes recombinantly in bacteria and characterized their biochemical properties. Whereas the overall genomic organization of Physarum NOS genes is comparable with various animal NOSs, none of the exon–intron boundaries are conserved. Recombinant expression of clones with various N-termini identified N-terminal amino acids essential for enzyme activity, but not required for haem binding or dimerization, and suggests the usage of non-AUG start codons for Physarum NOSs. Biochemical characterization of the two Physarum isoenzymes revealed different affinities for L-arginine, FMN and 6R-5,6,7,8-tetrahydro-L-biopterin. Portland Press Ltd. 2009-02-25 2009-03-15 /pmc/articles/PMC2677215/ /pubmed/19046139 http://dx.doi.org/10.1042/BJ20080192 Text en © 2009 The Author(s) The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by-nc/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Messner, Simon
Leitner, Stephan
Bommassar, Christian
Golderer, Georg
Gröbner, Peter
Werner, Ernst R.
Werner-Felmayer, Gabriele
Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title_full Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title_fullStr Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title_full_unstemmed Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title_short Physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
title_sort physarum nitric oxide synthases: genomic structures and enzymology of recombinant proteins
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2677215/
https://www.ncbi.nlm.nih.gov/pubmed/19046139
http://dx.doi.org/10.1042/BJ20080192
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