Oxygenase Domain of Drosophila melanogaster Nitric Oxide Synthase: Unique Kinetic Parameters Enable a More Efficient NO Release

Ray, S. S., Tejero, J., Wang, Z. Q., Dutta, T., Bhattacharjee, A., Regulski, M. R., Tully, T., Ghosh, S., Stuehr, D. J. (2007) Oxygenase Domain of Drosophila melanogaster Nitric Oxide Synthase: Unique Kinetic Parameters Enable a More Efficient NO Release. Biochemistry, 46 (42). pp. 11857-11864. ISSN 0006-2960 (Print)

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URL: http://pubs.acs.org/cgi-bin/article.cgi/bichaw/200...
DOI: 10.1021/bi700803p S0006-2960(70)00803-6

Abstract

Although nitric oxide (NO) is important for cell signaling and nonspecific immunity in the fruit fly Drosophila melanogaster, little is known about its single NO synthase (dNOS). We expressed the oxygenase domain of dNOS (dNOSoxy), characterized its spectroscopic, kinetic, and catalytic properties, and interpreted them in light of a global kinetic model for NO synthesis. Single turnover reactions with ferrous dNOSoxy showed it could convert Arg to N'omega-hydroxy-l-arginine (NOHA), or NOHA to citrulline and NO, when it was given 6R-tetrahydrobiopterin and O2. The dNOSoxy catalyzed Arg hydroxylation and NOHA oxidation at rates that matched or exceeded the rates catalyzed by the three mammalian NOSoxy enzymes. Consecutive heme-dioxy, ferric heme-NO, and ferric heme species were observed in the NOHA reaction of dNOSoxy, indicating that its catalytic mechanism is the same as in the mammalian NOS. However, NO dissociation from dNOSoxy was 4 to 9 times faster than that from the mammalian NOS enzymes. In contrast, the dNOSoxy ferrous heme-NO complex was relatively unreactive toward O2 and in this way was equivalent to the mammalian neuronal NOS. Our data show that dNOSoxy has unique settings for the kinetic parameters that determine its NO synthesis. Computer simulations reveal that these unique settings should enable dNOS to be a more efficient and active NO synthase than the mammalian NOS enzymes, which may allow it to function more broadly in cell signaling and immune functions in the fruit fly.

Item Type: Paper
Uncontrolled Keywords: TETRAHYDROBIOPTERIN RADICAL FORMATION HEME-DIOXY REDUCTION Arginine Hydroxylation CATALYTIC PROFILES BIDOMAIN STRUCTURE BACILLUS-SUBTILIS LOW-TEMPERATURE BINDING CALMODULIN EXPRESSION
Subjects: organism description > animal > insect > Drosophila
organism description > animal > mammal
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > nitric oxide synthase
CSHL Authors:
Depositing User: CSHL Librarian
Date Deposited: 03 Nov 2011 15:31
Last Modified: 29 Nov 2011 20:08
URI: http://repository.cshl.edu/id/eprint/23127

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