Eur J Biochem. 1996 Jul 15;239(2):418-26.

L-aspartate oxidase from Escherichia coli. I. Characterization of coenzyme binding and product inhibition.

Mortarino M, Negri A, Tedeschi G, Simonic T, Duga S, Gassen HG, Ronchi S.

Source

Istituto di Fisiologia Veterinaria e Biochimica, Università di Milano, Italy.

Abstract

This paper reports the biochemical characterization of the flavoprotein L-aspartate oxidase from Escherichia coli. Modification of a previously published procedure allowed overexpression of the holoenzyme in an unproteolysed form. L-Aspartate oxidase is a monomer of 60 kDa containing 1 mol of noncovalently bound FAD/mol protein. A polarographic and two spectrophotometric coupled assays have been set up to monitor the enzymatic activity continuously. L-Aspartate oxidase was subjected to product inhibition since iminoaspartate, which results from the oxidation of L-aspartate, binds to the enzyme with a dissociation constant (Kd) equal to 1.4 microM. The enzyme binds FAD by a simple second-order process with Kd 0.67 microM. Site-directed mutagenesis of the residues E43, G44, S45, F47 and Y48 located in the putative binding site of the isoallossazinic portion of FAD reduces the affinity for the coenzyme.

PMID:: 8706749; [PubMed - indexed for MEDLINE]

Free full text

Publication Types, MeSH Terms, Substances

Publication Types

MeSH Terms

Substances

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Covalent flavinylation of L-aspartate oxidase from Escherichia coli using N6-(6-carboxyhexyl)-FAD succinimidoester. [J Protein Chem. 1999]
Covalent flavinylation of L-aspartate oxidase from Escherichia coli using N6-(6-carboxyhexyl)-FAD succinimidoester.
Negri A, Buckmann AF, Tedeschi G, Stocker A, Ceciliani F, Treu C, Ronchi S. J Protein Chem. 1999 Aug; 18(6):671-6.
Mutagenesis at a highly conserved tyrosine in monoamine oxidase B affects FAD incorporation and catalytic activity. [Biochemistry. 1995]
Mutagenesis at a highly conserved tyrosine in monoamine oxidase B affects FAD incorporation and catalytic activity.
Zhou BP, Lewis DA, Kwan SW, Kirksey TJ, Abell CW. Biochemistry. 1995 Jul 25; 34(29):9526-31.
Structural characterization of l-aspartate oxidase and identification of an interdomain loop by limited proteolysis. [Eur J Biochem. 1999]
Structural characterization of l-aspartate oxidase and identification of an interdomain loop by limited proteolysis.
Tedeschi G, Negri A, Ceciliani F, Mattevi A, Ronchi S. Eur J Biochem. 1999 Mar; 260(3):896-903.
Redox potentials and quinone reductase activity of L-aspartate oxidase from Escherichia coli. [Biochemistry. 1997]
Redox potentials and quinone reductase activity of L-aspartate oxidase from Escherichia coli.
Tedeschi G, Zetta L, Negri A, Mortarino M, Ceciliani F, Ronchi S. Biochemistry. 1997 Dec 23; 36(51):16221-30.
Review A novel oxidoreductase family sharing a conserved FAD-binding domain. [Trends Biochem Sci. 1998]
Review A novel oxidoreductase family sharing a conserved FAD-binding domain.
Fraaije MW, Van Berkel WJ, Benen JA, Visser J, Mattevi A. Trends Biochem Sci. 1998 Jun; 23(6):206-7.

See reviews... See all...

Cited by 3 PubMed Central articles

Two sources of endogenous hydrogen peroxide in Escherichia coli. [Mol Microbiol. 2010]
Two sources of endogenous hydrogen peroxide in Escherichia coli.
Korshunov S, Imlay JA. Mol Microbiol. 2010 Mar; 75(6):1389-401. Epub 2010 Feb 8.
Characterization of quinolinate synthases from Escherichia coli, Mycobacterium tuberculosis, and Pyrococcus horikoshii indicates that [4Fe-4S] clusters are common cofactors throughout this class of enzymes. [Biochemistry. 2008]
Characterization of quinolinate synthases from Escherichia coli, Mycobacterium tuberculosis, and Pyrococcus horikoshii indicates that [4Fe-4S] clusters are common cofactors throughout this class of enzymes.
Saunders AH, Griffiths AE, Lee KH, Cicchillo RM, Tu L, Stromberg JA, Krebs C, Booker SJ. Biochemistry. 2008 Oct 14; 47(41):10999-1012. Epub 2008 Sep 20.
Review Linkage map of Escherichia coli K-12, edition 10: the traditional map. [Microbiol Mol Biol Rev. 1998]
Review Linkage map of Escherichia coli K-12, edition 10: the traditional map.
Berlyn MK. Microbiol Mol Biol Rev. 1998 Sep; 62(3):814-984.

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
BioSystems
Pathways and biological systems (BioSystems) that cite the current articles. Citations are from the BioSystems source databases (KEGG and BioCyc).
Compound (MeSH Keyword)
PubChem chemical compound records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Gene
Gene records that cite the current articles. Citations in Gene are added manually by NCBI or imported from outside public resources.
Nucleotide (Weighted)
Nucleotide records associated with the current articles through the Gene database. These are the related sequences on the Gene record that are added manually by NCBI.
Protein (RefSeq)
NCBI protein Reference Sequences (RefSeqs) that are cited in the current articles, included in the corresponding Gene Reference into Function, or that include the PubMed articles as references.
Protein (Weighted)
Protein records associated with the current articles through related Gene database records. These are the related sequences on the Gene record that are added manually by NCBI.
Protein Clusters
Clusters of related proteins from the Protein Clusters database that cite the current articles. Sources of references in Protein Clusters include the associated Gene and Conserved Domain records as well as NCBI added citations.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Taxonomy via GenBank
Taxonomy records associated with the current articles through taxonomic information on related molecular database records (Nucleotide, Protein, Gene, SNP, Structure).
GEO Profiles
Gene Expression Omnibus (GEO) Profiles of molecular abundance data. The current articles are references on the Gene record associated with the GEO profile.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

L-aspartate oxidase from Escherichia coli. I. Characterization of coenzyme bindi...
L-aspartate oxidase from Escherichia coli. I. Characterization of coenzyme binding and product inhibition.
Eur J Biochem. 1996 Jul 15 ;239(2):418-26.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: