NADB_ECOLI
ID NADB_ECOLI Reviewed; 540 AA.
AC P10902; P78099; Q2MAF6;
DT 01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT 29-AUG-2003, sequence version 4.
DT 03-AUG-2022, entry version 186.
DE RecName: Full=L-aspartate oxidase {ECO:0000303|PubMed:7033218};
DE Short=LASPO {ECO:0000303|PubMed:10425677};
DE EC=1.4.3.16 {ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:12200425, ECO:0000269|PubMed:20149100, ECO:0000269|PubMed:2187483, ECO:0000269|PubMed:2841129, ECO:0000269|PubMed:7033218, ECO:0000269|PubMed:8706749, ECO:0000269|PubMed:8706750};
DE AltName: Full=L-aspartate:fumarate oxidoreductase {ECO:0000303|PubMed:8706750};
DE EC=1.5.99.- {ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:12200425, ECO:0000269|PubMed:20149100, ECO:0000269|PubMed:8706750};
DE AltName: Full=Quinolinate synthetase B {ECO:0000303|PubMed:2841129};
GN Name=nadB {ECO:0000303|PubMed:2841129}; Synonyms=nicB;
GN OrderedLocusNames=b2574, JW2558;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-19, FUNCTION,
RP CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=2841129; DOI=10.1111/j.1432-1033.1988.tb14187.x;
RA Flachmann R., Kunz N., Seifert J., Guetlich M., Wientjes F.-J., Laeufer A.,
RA Gassen H.G.;
RT "Molecular biology of pyridine nucleotide biosynthesis in Escherichia coli.
RT Cloning and characterization of quinolinate synthesis genes nadA and
RT nadB.";
RL Eur. J. Biochem. 175:221-228(1988).
RN [2]
RP SEQUENCE REVISION.
RA Kunz N.;
RL Submitted (APR-1989) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RA Nashimoto H.;
RT "Non-ribosomal proteins affecting the assembly of ribosomes in Escherichia
RT coli.";
RL (In) Nierhaus K.H. (eds.);
RL The translational apparatus, pp.185-195, Plenum Press, New York (1993).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RA Nashimoto H., Saito N.;
RL Submitted (SEP-1995) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [6]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [7]
RP PARTIAL PROTEIN SEQUENCE, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=2187483;
RA Seifert J., Kunz N., Flachmann R., Laeufer A., Jany K.-D., Gassen H.G.;
RT "Expression of the E. coli nadB gene and characterization of the gene
RT product L-aspartate oxidase.";
RL Biol. Chem. Hoppe-Seyler 371:239-248(1990).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=7033218; DOI=10.1016/s0021-9258(19)68239-6;
RA Nasu S., Wicks F.D., Gholson R.K.;
RT "L-Aspartate oxidase, a newly discovered enzyme of Escherichia coli, is the
RT B protein of quinolinate synthetase.";
RL J. Biol. Chem. 257:626-632(1982).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, SUBUNIT, AND
RP MUTAGENESIS OF 43-GLU--SER-45; 43-GLU--TYR-48; GLU-43 AND SER-45.
RX PubMed=8706749; DOI=10.1111/j.1432-1033.1996.0418u.x;
RA Mortarino M., Negri A., Tedeschi G., Simonic T., Duga S., Gassen H.G.,
RA Ronchi S.;
RT "L-aspartate oxidase from Escherichia coli. I. Characterization of coenzyme
RT binding and product inhibition.";
RL Eur. J. Biochem. 239:418-426(1996).
RN [10]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8706750; DOI=10.1111/j.1432-1033.1996.0427u.x;
RA Tedeschi G., Negri A., Mortarino M., Ceciliani F., Simonic T., Faotto L.,
RA Ronchi S.;
RT "L-aspartate oxidase from Escherichia coli. II. Interaction with C4
RT dicarboxylic acids and identification of a novel L-aspartate: fumarate
RT oxidoreductase activity.";
RL Eur. J. Biochem. 239:427-433(1996).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP MUTAGENESIS OF HIS-244; ARG-290; HIS-351 AND ARG-386, AND ACTIVE SITE.
RX PubMed=11294641; DOI=10.1021/bi002406u;
RA Tedeschi G., Ronchi S., Simonic T., Treu C., Mattevi A., Negri A.;
RT "Probing the active site of L-aspartate oxidase by site-directed
RT mutagenesis: role of basic residues in fumarate reduction.";
RL Biochemistry 40:4738-4744(2001).
RN [12]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=12200425; DOI=10.1074/jbc.m204958200;
RA Messner K.R., Imlay J.A.;
RT "Mechanism of superoxide and hydrogen peroxide formation by fumarate
RT reductase, succinate dehydrogenase, and aspartate oxidase.";
RL J. Biol. Chem. 277:42563-42571(2002).
RN [13]
RP FUNCTION, CATALYTIC ACTIVITY, AND DISRUPTION PHENOTYPE.
RX PubMed=20149100; DOI=10.1111/j.1365-2958.2010.07059.x;
RA Korshunov S., Imlay J.A.;
RT "Two sources of endogenous hydrogen peroxide in Escherichia coli.";
RL Mol. Microbiol. 75:1389-1401(2010).
RN [14]
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF GLU-121.
RX PubMed=20600565; DOI=10.1016/j.biochi.2010.06.015;
RA Tedeschi G., Nonnis S., Strumbo B., Cruciani G., Carosati E., Negri A.;
RT "On the catalytic role of the active site residue E121 of E. coli L-
RT aspartate oxidase.";
RL Biochimie 92:1335-1342(2010).
RN [15]
RP REACTION MECHANISM.
RX PubMed=28700220; DOI=10.1021/acs.biochem.7b00307;
RA Chow C., Hegde S., Blanchard J.S.;
RT "Mechanistic characterization of Escherichia coli L-aspartate oxidase from
RT kinetic isotope effects.";
RL Biochemistry 56:4044-4052(2017).
RN [16] {ECO:0007744|PDB:1CHU}
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS), SUBUNIT, AND DOMAIN.
RX PubMed=10425677; DOI=10.1016/s0969-2126(99)80099-9;
RA Mattevi A., Tedeschi G., Bacchella L., Coda A., Negri A., Ronchi S.;
RT "Structure of L-aspartate oxidase: implications for the succinate
RT dehydrogenase/fumarate reductase oxidoreductase family.";
RL Structure 7:745-756(1999).
RN [17] {ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) OF MUTANT ILE-386 IN COMPLEX WITH
RP FAD AND SUCCINATE, COFACTOR, DOMAIN, AND ACTIVE SITE.
RX PubMed=11863440; DOI=10.1021/bi015939r;
RA Bossi R.T., Negri A., Tedeschi G., Mattevi A.;
RT "Structure of FAD-bound L-aspartate oxidase: insight into substrate
RT specificity and catalysis.";
RL Biochemistry 41:3018-3024(2002).
CC -!- FUNCTION: Catalyzes the oxidation of L-aspartate to iminoaspartate, the
CC first step in the de novo biosynthesis of NAD(+) (PubMed:2841129,
CC PubMed:7033218, PubMed:2187483, PubMed:8706749, PubMed:8706750,
CC PubMed:11294641). Can use either oxygen or fumarate as electron
CC acceptors, which allows the enzyme to be functional under aerobic and
CC anaerobic conditions (PubMed:8706750, PubMed:11294641,
CC PubMed:12200425). In vivo, fumarate is used under anaerobic conditions,
CC and oxygen is the predominant electron acceptor under aerobic
CC conditions due to the lower fumarate levels (PubMed:20149100). In
CC vitro, fumarate is a more efficient electron acceptor and is
CC kinetically superior to oxygen (PubMed:12200425, PubMed:20149100).
CC {ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:12200425,
CC ECO:0000269|PubMed:20149100, ECO:0000269|PubMed:2187483,
CC ECO:0000269|PubMed:2841129, ECO:0000269|PubMed:7033218,
CC ECO:0000269|PubMed:8706749, ECO:0000269|PubMed:8706750}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-aspartate + O2 = H2O2 + iminosuccinate;
CC Xref=Rhea:RHEA:25876, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:29991, ChEBI:CHEBI:77875; EC=1.4.3.16;
CC Evidence={ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:12200425,
CC ECO:0000269|PubMed:20149100, ECO:0000269|PubMed:2187483,
CC ECO:0000269|PubMed:2841129, ECO:0000269|PubMed:7033218,
CC ECO:0000269|PubMed:8706749, ECO:0000269|PubMed:8706750};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25877;
CC Evidence={ECO:0000269|PubMed:2841129, ECO:0000269|PubMed:7033218,
CC ECO:0000269|PubMed:8706750};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=fumarate + L-aspartate = iminosuccinate + succinate;
CC Xref=Rhea:RHEA:30043, ChEBI:CHEBI:29806, ChEBI:CHEBI:29991,
CC ChEBI:CHEBI:30031, ChEBI:CHEBI:77875;
CC Evidence={ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:12200425,
CC ECO:0000269|PubMed:20149100, ECO:0000269|PubMed:8706750};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:30044;
CC Evidence={ECO:0000269|PubMed:8706750};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:11863440,
CC ECO:0000269|PubMed:2187483, ECO:0000269|PubMed:7033218,
CC ECO:0000269|PubMed:8706749};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11863440};
CC -!- ACTIVITY REGULATION: Inhibited by the product iminoaspartate
CC (PubMed:8706749). Competitively inhibited by mesotartrate
CC (PubMed:2187483). NAD acts as a competitive inhibitor to FAD
CC (PubMed:7033218, PubMed:2187483). Inhibited by iodoacetic acid,
CC diethylpyrocarbonate and tetranitromethane (PubMed:2187483).
CC {ECO:0000269|PubMed:2187483, ECO:0000269|PubMed:7033218,
CC ECO:0000269|PubMed:8706749}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.63 mM for L-aspartate (at substrate concentrations between 0.25
CC and 1.25 mM) {ECO:0000269|PubMed:7033218};
CC KM=3.33 mM for L-aspartate (at substrate concentrations between 2.0
CC and 10.00 mM) {ECO:0000269|PubMed:7033218};
CC KM=0.5 mM for L-aspartate (at substrate concentrations between 0.2
CC and l mM) {ECO:0000269|PubMed:2187483};
CC KM=4.1 mM for L-aspartate (at substrate concentrations between 1 and
CC 20 mM) {ECO:0000269|PubMed:2187483};
CC KM=4.9 mM for L-aspartate {ECO:0000269|PubMed:20600565};
CC KM=17.0 mM for N-acetyl-L-aspartate {ECO:0000269|PubMed:20600565};
CC KM=3.7 mM for N-formyl-L-aspartate {ECO:0000269|PubMed:20600565};
CC KM=4.7 mM for 3-OH-erythro-L-aspartate {ECO:0000269|PubMed:20600565};
CC KM=2.5 mM for FAD {ECO:0000269|PubMed:7033218};
CC Note=kcat is 29.4 min(-1) for L-aspartate oxidase activity with
CC oxygen as electron acceptor. kcat is 16.0 min(-1) for reduced benzyl
CC viologen:fumarate oxidoreductase activity (PubMed:11294641). kcat is
CC 40.0 min(-1) with L-aspartate as substrate. kcat is 0.21 min(-1) with
CC N-acetyl-L-aspartate as substrate. kcat is 0.46 min(-1) with N-
CC formyl-L-aspartate as substrate. kcat is 10.4 min(-1) with 3-OH-
CC erythro-L-aspartate as substrate (PubMed:20600565).
CC {ECO:0000269|PubMed:11294641, ECO:0000269|PubMed:20600565};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:7033218};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:2187483};
CC -!- PATHWAY: Cofactor biosynthesis; NAD(+) biosynthesis; iminoaspartate
CC from L-aspartate (oxidase route): step 1/1.
CC {ECO:0000269|PubMed:7033218, ECO:0000305|PubMed:2841129}.
CC -!- SUBUNIT: Monomer (PubMed:2187483, PubMed:8706749). Homodimer
CC (PubMed:10425677). Both the monomeric and dimeric forms of the enzyme
CC are catalytically active (PubMed:10425677).
CC {ECO:0000269|PubMed:10425677, ECO:0000269|PubMed:2187483,
CC ECO:0000269|PubMed:8706749}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- DOMAIN: Folds into three domains: the FAD-binding domain, the capping
CC domain and the C-terminal helical domain. {ECO:0000269|PubMed:10425677,
CC ECO:0000269|PubMed:11863440}.
CC -!- DISRUPTION PHENOTYPE: Mutants lacking this gene generate 25% less
CC H(2)O(2) than the parent strain. {ECO:0000269|PubMed:20149100}.
CC -!- MISCELLANEOUS: The chemistry is similar to that of typical amino acid
CC oxidases in which the transfer of the hydride from C2 of L-aspartate to
CC FAD is rate-limiting and occurs in a concerted manner with respect to
CC deprotonation of the alpha-amine. NadB may have structurally evolved
CC from succinate dehydrogenase/fumarate reductase-type enzymes to gain
CC the new functionality of oxidizing amino acids while retaining the
CC ability to reduce fumarate. {ECO:0000269|PubMed:28700220}.
CC -!- SIMILARITY: Belongs to the FAD-dependent oxidoreductase 2 family. NadB
CC subfamily. {ECO:0000305}.
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DR EMBL; X12714; CAA31217.1; -; Genomic_DNA.
DR EMBL; D13169; BAA02446.1; -; Genomic_DNA.
DR EMBL; D64044; BAA10921.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75627.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76750.1; -; Genomic_DNA.
DR PIR; E65035; OXECLD.
DR RefSeq; NP_417069.1; NC_000913.3.
DR RefSeq; WP_001094491.1; NZ_LN832404.1.
DR PDB; 1CHU; X-ray; 2.20 A; A=1-540.
DR PDB; 1KNP; X-ray; 2.60 A; A=1-540.
DR PDB; 1KNR; X-ray; 2.50 A; A=1-540.
DR PDBsum; 1CHU; -.
DR PDBsum; 1KNP; -.
DR PDBsum; 1KNR; -.
DR AlphaFoldDB; P10902; -.
DR SMR; P10902; -.
DR BioGRID; 4263282; 13.
DR BioGRID; 851387; 1.
DR DIP; DIP-556N; -.
DR IntAct; P10902; 7.
DR STRING; 511145.b2574; -.
DR DrugBank; DB03147; Flavin adenine dinucleotide.
DR PaxDb; P10902; -.
DR PRIDE; P10902; -.
DR EnsemblBacteria; AAC75627; AAC75627; b2574.
DR EnsemblBacteria; BAE76750; BAE76750; BAE76750.
DR GeneID; 947049; -.
DR KEGG; ecj:JW2558; -.
DR KEGG; eco:b2574; -.
DR PATRIC; fig|1411691.4.peg.4160; -.
DR EchoBASE; EB0625; -.
DR eggNOG; COG0029; Bacteria.
DR HOGENOM; CLU_014312_3_0_6; -.
DR InParanoid; P10902; -.
DR OMA; HCVQWLI; -.
DR PhylomeDB; P10902; -.
DR BioCyc; EcoCyc:L-ASPARTATE-OXID-MON; -.
DR BioCyc; MetaCyc:L-ASPARTATE-OXID-MON; -.
DR BRENDA; 1.4.3.16; 2026.
DR UniPathway; UPA00253; UER00326.
DR EvolutionaryTrace; P10902; -.
DR PRO; PR:P10902; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IDA:EcoCyc.
DR GO; GO:0008734; F:L-aspartate oxidase activity; IDA:EcoCyc.
DR GO; GO:0044318; F:L-aspartate:fumarate oxidoreductase activity; IDA:EcoCyc.
DR GO; GO:0034628; P:'de novo' NAD biosynthetic process from aspartate; IMP:EcoCyc.
DR Gene3D; 3.50.50.60; -; 1.
DR Gene3D; 3.90.700.10; -; 1.
DR InterPro; IPR003953; FAD-binding_2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR037099; Fum_R/Succ_DH_flav-like_C_sf.
DR InterPro; IPR015939; Fum_Rdtase/Succ_DH_flav-like_C.
DR InterPro; IPR005288; NadB.
DR InterPro; IPR027477; Succ_DH/fumarate_Rdtase_cat_sf.
DR PANTHER; PTHR42716; PTHR42716; 1.
DR Pfam; PF00890; FAD_binding_2; 1.
DR Pfam; PF02910; Succ_DH_flav_C; 1.
DR SUPFAM; SSF46977; SSF46977; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR SUPFAM; SSF56425; SSF56425; 1.
DR TIGRFAMs; TIGR00551; nadB; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; FAD; Flavoprotein;
KW Nucleotide-binding; Oxidoreductase; Pyridine nucleotide biosynthesis;
KW Reference proteome.
FT CHAIN 1..540
FT /note="L-aspartate oxidase"
FT /id="PRO_0000184384"
FT ACT_SITE 290
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000305|PubMed:11294641,
FT ECO:0000305|PubMed:11863440, ECO:0007744|PDB:1KNP"
FT BINDING 16..19
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 38
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 45..52
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 161..162
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 223
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 244
FT /ligand="succinate"
FT /ligand_id="ChEBI:CHEBI:30031"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP"
FT BINDING 259..260
FT /ligand="succinate"
FT /ligand_id="ChEBI:CHEBI:30031"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP"
FT BINDING 375
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT BINDING 389
FT /ligand="succinate"
FT /ligand_id="ChEBI:CHEBI:30031"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP"
FT BINDING 391..392
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:11863440,
FT ECO:0007744|PDB:1KNP, ECO:0007744|PDB:1KNR"
FT SITE 121
FT /note="Important in orienting the L-aspartate substrate"
FT /evidence="ECO:0000305|PubMed:11294641"
FT MUTAGEN 43..48
FT /note="EGSTFY->RSHTVA: Loss of activity. Has largely lost
FT both FAD and ligand-binding properties."
FT /evidence="ECO:0000269|PubMed:8706749"
FT MUTAGEN 43..45
FT /note="EGS->RSH: Loss of activity. Has largely lost both
FT FAD and ligand-binding properties."
FT /evidence="ECO:0000269|PubMed:8706749"
FT MUTAGEN 43
FT /note="E->R: 2-fold increase in Kd for FAD. Retains 15% of
FT specific activity."
FT /evidence="ECO:0000269|PubMed:8706749"
FT MUTAGEN 45
FT /note="S->H: Loss of activity. Has largely lost both FAD
FT and ligand-binding properties."
FT /evidence="ECO:0000269|PubMed:8706749"
FT MUTAGEN 121
FT /note="E->A: Retains reduced benzyl viologen:fumarate
FT oxidoreductase activity, with almost the same catalytic
FT efficiency. Lacks L-aspartate:oxygen and L-
FT aspartate:fumarate oxidoreductase activities."
FT /evidence="ECO:0000269|PubMed:20600565"
FT MUTAGEN 121
FT /note="E->D: Retains reduced benzyl viologen:fumarate
FT oxidoreductase activity, with 690-fold decrease in
FT catalytic efficiency. Lacks L-aspartate:oxygen and L-
FT aspartate:fumarate oxidoreductase activities."
FT /evidence="ECO:0000269|PubMed:20600565"
FT MUTAGEN 121
FT /note="E->K: Retains reduced benzyl viologen:fumarate
FT oxidoreductase activity, with 72-fold decrease in catalytic
FT efficiency. Lacks L-aspartate:oxygen and L-
FT aspartate:fumarate oxidoreductase activities."
FT /evidence="ECO:0000269|PubMed:20600565"
FT MUTAGEN 121
FT /note="E->Q: Retains reduced benzyl viologen:fumarate
FT oxidoreductase activity, with 17-fold decrease in catalytic
FT efficiency. Lacks L-aspartate:oxygen and L-
FT aspartate:fumarate oxidoreductase activities."
FT /evidence="ECO:0000269|PubMed:20600565"
FT MUTAGEN 244
FT /note="H->A: 18-fold decrease in catalytic efficiency with
FT both oxygen and fumarate as electron acceptors. Decreases
FT FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT MUTAGEN 244
FT /note="H->S: 122-fold decrease in catalytic efficiency with
FT oxygen as electron acceptor. 110-fold decrease in catalytic
FT efficiency with fumarate as electron acceptor. Decreases
FT FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT MUTAGEN 290
FT /note="R->L: Loss of activity with both oxygen and fumarate
FT as electron acceptors. Decreases FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT MUTAGEN 351
FT /note="H->A: 1170-fold decrease in catalytic efficiency
FT with oxygen as electron acceptor. 83-fold decrease in
FT catalytic efficiency with fumarate as electron acceptor.
FT Decreases FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT MUTAGEN 351
FT /note="H->S: Shows weak activity with oxygen as electron
FT acceptor. 148-fold decrease in catalytic efficiency with
FT fumarate as electron acceptor. Decreases FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT MUTAGEN 386
FT /note="R->L: Shows weak activity with oxygen as electron
FT acceptor. 78-fold decrease in catalytic efficiency with
FT fumarate as electron acceptor. Decreases FAD binding."
FT /evidence="ECO:0000269|PubMed:11294641"
FT CONFLICT 160
FT /note="S -> T (in Ref. 1, 3 and 4)"
FT /evidence="ECO:0000305"
FT CONFLICT 485
FT /note="N -> D (in Ref. 1; CAA31217)"
FT /evidence="ECO:0000305"
FT STRAND 6..8
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 10..14
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 18..27
FT /evidence="ECO:0007829|PDB:1CHU"
FT TURN 28..30
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 33..36
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 38..40
FT /evidence="ECO:0007829|PDB:1KNR"
FT HELIX 45..48
FT /evidence="ECO:0007829|PDB:1KNR"
FT HELIX 60..73
FT /evidence="ECO:0007829|PDB:1CHU"
FT TURN 74..76
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 80..99
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 110..113
FT /evidence="ECO:0007829|PDB:1KNR"
FT HELIX 145..150
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 154..157
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 159..167
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 168..170
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 178..186
FT /evidence="ECO:0007829|PDB:1CHU"
FT TURN 187..190
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 191..196
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 198..202
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 208..210
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 211..215
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 217..219
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 223..230
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 241..248
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 260..264
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 268..270
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 278..280
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 285..287
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 290..304
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 309..312
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 319..324
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 326..333
FT /evidence="ECO:0007829|PDB:1CHU"
FT TURN 334..336
FT /evidence="ECO:0007829|PDB:1CHU"
FT TURN 339..341
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 344..354
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 356..358
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 366..368
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 370..372
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 374..376
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 380..382
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 390..409
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 410..412
FT /evidence="ECO:0007829|PDB:1KNR"
FT HELIX 430..450
FT /evidence="ECO:0007829|PDB:1CHU"
FT STRAND 451..455
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 457..478
FT /evidence="ECO:0007829|PDB:1CHU"
FT HELIX 485..506
FT /evidence="ECO:0007829|PDB:1CHU"
SQ SEQUENCE 540 AA; 60337 MW; 7B6CFF633BD4AFEC CRC64;
MNTLPEHSCD VLIIGSGAAG LSLALRLADQ HQVIVLSKGP VTEGSTFYAQ GGIAAVFDET
DSIDSHVEDT LIAGAGICDR HAVEFVASNA RSCVQWLIDQ GVLFDTHIQP NGEESYHLTR
EGGHSHRRIL HAADATGREV ETTLVSKALN HPNIRVLERS NAVDLIVSDK IGLPGTRRVV
GAWVWNRNKE TVETCHAKAV VLATGGASKV YQYTTNPDIS SGDGIAMAWR AGCRVANLEF
NQFHPTALYH PQARNFLLTE ALRGEGAYLK RPDGTRFMPD FDERGELAPR DIVARAIDHE
MKRLGADCMF LDISHKPADF IRQHFPMIYE KLLGLGIDLT QEPVPIVPAA HYTCGGVMVD
DHGRTDVEGL YAIGEVSYTG LHGANRMASN SLLECLVYGW SAAEDITRRM PYAHDISTLP
PWDESRVENP DERVVIQHNW HELRLFMWDY VGIVRTTKRL ERALRRITML QQEIDEYYAH
FRVSNNLLEL RNLVQVAELI VRCAMMRKES RGLHFTLDYP ELLTHSGPSI LSPGNHYINR
