HLNO_PAENI
ID HLNO_PAENI Reviewed; 425 AA.
AC Q93NH4; O86101;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 03-AUG-2022, entry version 110.
DE RecName: Full=(S)-6-hydroxynicotine oxidase {ECO:0000305};
DE EC=1.5.3.5 {ECO:0000269|PubMed:21383134, ECO:0000269|PubMed:26744768, ECO:0000269|PubMed:28080034, ECO:0000269|PubMed:4965794, ECO:0000269|PubMed:5646150, ECO:0000269|PubMed:5849820};
DE AltName: Full=6-hydroxy-L-nicotine oxidase {ECO:0000303|PubMed:9878353};
DE Short=6-HLNO {ECO:0000303|PubMed:9878353};
DE AltName: Full=L-6-hydroxynicotine oxidase {ECO:0000303|PubMed:4965794};
DE AltName: Full=L-hydroxynicotine oxidase {ECO:0000303|PubMed:26744768};
DE Short=LHNO {ECO:0000303|PubMed:26744768};
GN Name=6-hlno {ECO:0000312|EMBL:CAD47951.1};
OS Paenarthrobacter nicotinovorans (Arthrobacter nicotinovorans).
OG Plasmid pAO1.
OC Bacteria; Actinobacteria; Micrococcales; Micrococcaceae; Paenarthrobacter.
OX NCBI_TaxID=29320;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 49919 / DSM 420 / JCM 3874 / NBRC 15511; PLASMID=pAO1;
RX PubMed=9878353; DOI=10.1006/jmbi.1998.2227;
RA Schenk S., Hoelz A., Kraus B., Decker K.;
RT "Gene structures and properties of enzymes of the plasmid-encoded nicotine
RT catabolism of Arthrobacter nicotinovorans.";
RL J. Mol. Biol. 284:1323-1339(1998).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 49919 / DSM 420 / JCM 3874 / NBRC 15511; PLASMID=pAO1;
RX PubMed=9929386; DOI=10.1007/pl00006456;
RA Schenk S., Decker K.;
RT "Horizontal gene transfer involved in the convergent evolution of the
RT plasmid-encoded enantioselective 6-hydroxynicotine oxidases.";
RL J. Mol. Evol. 48:178-186(1999).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 49919 / DSM 420 / JCM 3874 / NBRC 15511; PLASMID=pAO1;
RX PubMed=11514508; DOI=10.1128/jb.183.18.5262-5267.2001;
RA Baitsch D., Sandu C., Brandsch R., Igloi G.L.;
RT "Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in
RT degradation of the plant alkaloid nicotine: cloning, purification, and
RT characterization of 2,6-dihydroxypyridine 3-hydroxylase.";
RL J. Bacteriol. 183:5262-5267(2001).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 49919 / DSM 420 / JCM 3874 / NBRC 15511; PLASMID=pAO1;
RX PubMed=12618462; DOI=10.1128/jb.185.6.1976-1986.2003;
RA Igloi G.L., Brandsch R.;
RT "Sequence of the 165-kilobase catabolic plasmid pAO1 from Arthrobacter
RT nicotinovorans and identification of a pAO1-dependent nicotine uptake
RT system.";
RL J. Bacteriol. 185:1976-1986(2003).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, PATHWAY, AND INDUCTION.
RX PubMed=5849820; DOI=10.1016/s0926-6593(65)80155-2;
RA Decker K., Bleeg H.;
RT "Induction and purification of stereospecific nicotine oxidizing enzymes
RT from Arthrobacter oxidans.";
RL Biochim. Biophys. Acta 105:313-324(1965).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND ACTIVITY REGULATION.
RX PubMed=4965794; DOI=10.1111/j.1432-1033.1967.tb19507.x;
RA Decker K., Dai V.D.;
RT "Mechanism and specifcity of L- and D-6-hydroxynicotine oxidase.";
RL Eur. J. Biochem. 3:132-138(1967).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=5646150; DOI=10.1111/j.1432-1033.1968.tb00177.x;
RA Dai V.D., Decker K., Sund H.;
RT "Purification and properties of L-6-hydroxynicotine oxidase.";
RL Eur. J. Biochem. 4:95-102(1968).
RN [8]
RP SUBCELLULAR LOCATION, AND INDUCTION.
RX PubMed=4019415; DOI=10.1128/jb.163.2.792-795.1985;
RA Swafford J.R., Reeves H.C., Brandsch R.;
RT "Localization of the enantiozymes of 6-hydroxy-nicotine oxidase in
RT Arthrobacter oxidans by electron immunochemistry.";
RL J. Bacteriol. 163:792-795(1985).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND MUTAGENESIS OF HIS-187; GLU-300 AND TYR-407.
RX PubMed=26744768; DOI=10.1021/acs.biochem.5b01325;
RA Fitzpatrick P.F., Chadegani F., Zhang S., Roberts K.M., Hinck C.S.;
RT "Mechanism of the flavoprotein L-hydroxynicotine oxidase: kinetic
RT mechanism, substrate specificity, reaction product, and roles of active-
RT site residues.";
RL Biochemistry 55:697-703(2016).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, AND MUTAGENESIS OF
RP ASN-166; LYS-287 AND TYR-311.
RX PubMed=28080034; DOI=10.1021/acs.biochem.6b01160;
RA Fitzpatrick P.F., Chadegani F., Zhang S., Dougherty V.;
RT "Mechanism of flavoprotein L-6-hydroxynicotine oxidase: pH and solvent
RT isotope effects and identification of key active site residues.";
RL Biochemistry 56:869-875(2017).
RN [11] {ECO:0007744|PDB:3K7M, ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEXES WITH FAD AND
RP 6-HYDROXY-L-NICOTINE, COFACTOR, SUBUNIT, AND DOMAIN.
RX PubMed=20006620; DOI=10.1016/j.jmb.2009.12.009;
RA Kachalova G.S., Bourenkov G.P., Mengesdorf T., Schenk S., Maun H.R.,
RA Burghammer M., Riekel C., Decker K., Bartunik H.D.;
RT "Crystal structure analysis of free and substrate-bound 6-hydroxy-L-
RT nicotine oxidase from Arthrobacter nicotinovorans.";
RL J. Mol. Biol. 396:785-799(2010).
RN [12] {ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC, ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO, ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1, ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0, ECO:0007744|PDB:3NN6}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) IN COMPLEXES WITH FAD;
RP 6-HYDROXY-L-NICOTINE; 6-HYDROXY-D-NICOTINE; 6-HYDROXY-N-METHYLMYOSMINE;
RP 6-HYDROXYPSEUDOOXYNICOTINE AND INHIBITORS, FUNCTION, CATALYTIC ACTIVITY,
RP COFACTOR, AND DOMAIN.
RX PubMed=21383134; DOI=10.1073/pnas.1016684108;
RA Kachalova G., Decker K., Holt A., Bartunik H.D.;
RT "Crystallographic snapshots of the complete reaction cycle of nicotine
RT degradation by an amine oxidase of the monoamine oxidase (MAO) family.";
RL Proc. Natl. Acad. Sci. U.S.A. 108:4800-4805(2011).
CC -!- FUNCTION: Involved in the degradation of L-nicotine (PubMed:5849820).
CC Catalyzes the oxidation of (S)-6-hydroxynicotine (6-hydroxy-L-nicotine)
CC to 6-hydroxypseudooxynicotine (PubMed:5849820, PubMed:4965794,
CC PubMed:5646150, PubMed:21383134, PubMed:26744768, PubMed:28080034).
CC Oxidation of the pyrrolidine ring of (S)-6-hydroxynicotine leads to the
CC formation of the optically inactive 6-hydroxy-N-methylmyosmine, which
CC hydrolyzes spontaneously to 6-hydroxypseudooxynicotine (PubMed:4965794,
CC PubMed:21383134, PubMed:26744768, PubMed:28080034). Acts with absolute
CC stereospecificity on the L-form of 6-hydroxynicotine (PubMed:4965794).
CC Can also use (S)-6-hydroxynornicotine (PubMed:26744768,
CC PubMed:28080034). {ECO:0000269|PubMed:21383134,
CC ECO:0000269|PubMed:26744768, ECO:0000269|PubMed:28080034,
CC ECO:0000269|PubMed:4965794, ECO:0000269|PubMed:5646150,
CC ECO:0000269|PubMed:5849820}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-6-hydroxynicotine + H2O + O2 = 6-hydroxypseudooxynicotine
CC + H2O2; Xref=Rhea:RHEA:11880, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:58182, ChEBI:CHEBI:58682; EC=1.5.3.5;
CC Evidence={ECO:0000269|PubMed:21383134, ECO:0000269|PubMed:26744768,
CC ECO:0000269|PubMed:28080034, ECO:0000269|PubMed:4965794,
CC ECO:0000269|PubMed:5646150, ECO:0000269|PubMed:5849820};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:11881;
CC Evidence={ECO:0000269|PubMed:21383134, ECO:0000269|PubMed:26744768,
CC ECO:0000269|PubMed:28080034, ECO:0000269|PubMed:4965794,
CC ECO:0000269|PubMed:5646150, ECO:0000269|PubMed:5849820};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-6-hydroxynicotine + O2 = 6-hydroxy-N-methylmyosmine +
CC H2O2; Xref=Rhea:RHEA:46976, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:58182, ChEBI:CHEBI:87164;
CC Evidence={ECO:0000269|PubMed:21383134, ECO:0000269|PubMed:26744768,
CC ECO:0000269|PubMed:28080034, ECO:0000269|PubMed:4965794};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:46977;
CC Evidence={ECO:0000269|PubMed:21383134, ECO:0000269|PubMed:26744768,
CC ECO:0000269|PubMed:28080034, ECO:0000269|PubMed:4965794};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:20006620, ECO:0000269|PubMed:21383134,
CC ECO:0000269|PubMed:4965794, ECO:0000269|PubMed:5646150};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:20006620,
CC ECO:0000269|PubMed:21383134};
CC -!- ACTIVITY REGULATION: Inhibited by (R)-6-hydroxynicotine
CC (PubMed:5849820, PubMed:4965794). Inhibited by high concentrations of
CC phenanthroline (PubMed:5646150). Activity is strongly affected by
CC Hg(2+) and p-chloromercuriphenylsulfonate, but not by N-ethylmaleimide
CC and 5,5'-dithiobis-(2-nitrobenzoate) (PubMed:5646150).
CC {ECO:0000269|PubMed:4965794, ECO:0000269|PubMed:5646150,
CC ECO:0000269|PubMed:5849820}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.02 mM for (S)-6-hydroxynicotine {ECO:0000269|PubMed:5849820};
CC KM=0.042 mM for (S)-6-hydroxynicotine {ECO:0000269|PubMed:26744768};
CC KM=0.064 mM for (S)-6-hydroxynornicotine
CC {ECO:0000269|PubMed:26744768};
CC Note=kcat is 30 sec(-1) with (S)-6-hydroxynicotine as substrate. kcat
CC is 16 sec(-1) with (S)-6-hydroxynornicotine as substrate.
CC {ECO:0000269|PubMed:26744768};
CC pH dependence:
CC Fairly stable between pH 6 and 9. {ECO:0000269|PubMed:5646150};
CC -!- PATHWAY: Alkaloid degradation; nicotine degradation; 6-
CC hydroxypseudooxynicotine from nicotine (S-isomer route): step 2/2.
CC {ECO:0000305|PubMed:5849820}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:20006620,
CC ECO:0000269|PubMed:5646150}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:4019415}.
CC -!- INDUCTION: Induced in the presence of L-nicotine and D-nicotine
CC (PubMed:5849820). Expressed during logarithmic growth and stationary
CC phase (PubMed:4019415). {ECO:0000269|PubMed:4019415,
CC ECO:0000269|PubMed:5849820}.
CC -!- DOMAIN: Consists of two domains, an FAD-binding domain and a substrate-
CC binding domain (PubMed:20006620). The inactive D-stereoisomer binds in
CC mirror symmetry with respect to the catalytic axis, revealing absolute
CC stereospecificity of hydrogen transfer to the flavin (PubMed:21383134).
CC {ECO:0000269|PubMed:20006620, ECO:0000269|PubMed:21383134}.
CC -!- SIMILARITY: Belongs to the flavin monoamine oxidase family.
CC {ECO:0000305}.
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DR EMBL; AJ223391; CAA11306.1; -; Genomic_DNA.
DR EMBL; AF373840; AAK64245.1; -; Genomic_DNA.
DR EMBL; AJ507836; CAD47951.1; -; Genomic_DNA.
DR RefSeq; WP_016359462.1; NC_021229.1.
DR RefSeq; YP_007988777.1; NC_021229.1.
DR PDB; 3K7M; X-ray; 1.95 A; X=1-425.
DR PDB; 3K7Q; X-ray; 2.05 A; X=1-425.
DR PDB; 3K7T; X-ray; 2.85 A; A/B=1-425.
DR PDB; 3NG7; X-ray; 1.95 A; X=1-425.
DR PDB; 3NGC; X-ray; 2.25 A; X=1-425.
DR PDB; 3NH3; X-ray; 2.10 A; X=1-425.
DR PDB; 3NHO; X-ray; 2.85 A; X=1-425.
DR PDB; 3NK0; X-ray; 2.15 A; X=1-425.
DR PDB; 3NK1; X-ray; 2.20 A; X=1-425.
DR PDB; 3NK2; X-ray; 2.65 A; X=1-425.
DR PDB; 3NN0; X-ray; 2.75 A; X=1-425.
DR PDB; 3NN6; X-ray; 2.19 A; X=1-425.
DR PDBsum; 3K7M; -.
DR PDBsum; 3K7Q; -.
DR PDBsum; 3K7T; -.
DR PDBsum; 3NG7; -.
DR PDBsum; 3NGC; -.
DR PDBsum; 3NH3; -.
DR PDBsum; 3NHO; -.
DR PDBsum; 3NK0; -.
DR PDBsum; 3NK1; -.
DR PDBsum; 3NK2; -.
DR PDBsum; 3NN0; -.
DR PDBsum; 3NN6; -.
DR SMR; Q93NH4; -.
DR KEGG; ag:CAA11306; -.
DR BioCyc; MetaCyc:MON-964; -.
DR BRENDA; 1.5.3.5; 449.
DR BRENDA; 1.5.99.14; 449.
DR BRENDA; 1.5.99.4; 449.
DR UniPathway; UPA00106; UER00919.
DR EvolutionaryTrace; Q93NH4; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0018531; F:(S)-6-hydroxynicotine oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0019608; P:nicotine catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR002937; Amino_oxidase.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR001613; Flavin_amine_oxidase.
DR Pfam; PF01593; Amino_oxidase; 1.
DR PRINTS; PR00757; AMINEOXDASEF.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Cytoplasm; FAD; Flavoprotein;
KW Nucleotide-binding; Oxidoreductase; Plasmid.
FT CHAIN 1..425
FT /note="(S)-6-hydroxynicotine oxidase"
FT /id="PRO_0000455450"
FT BINDING 12
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 31
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 38..39
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 56..59
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 166
FT /ligand="(S)-6-hydroxynicotine"
FT /ligand_id="ChEBI:CHEBI:58182"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7Q,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3"
FT BINDING 226
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 311
FT /ligand="(S)-6-hydroxynicotine"
FT /ligand_id="ChEBI:CHEBI:58182"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7Q,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3"
FT BINDING 326
FT /ligand="(S)-6-hydroxynicotine"
FT /ligand_id="ChEBI:CHEBI:58182"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7Q,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3"
FT BINDING 371
FT /ligand="(S)-6-hydroxynicotine"
FT /ligand_id="ChEBI:CHEBI:58182"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3NG7,
FT ECO:0007744|PDB:3NGC, ECO:0007744|PDB:3NH3"
FT BINDING 398
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 406..408
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7M,
FT ECO:0007744|PDB:3K7Q, ECO:0007744|PDB:3K7T,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3, ECO:0007744|PDB:3NHO,
FT ECO:0007744|PDB:3NK0, ECO:0007744|PDB:3NK1,
FT ECO:0007744|PDB:3NK2, ECO:0007744|PDB:3NN0"
FT BINDING 407
FT /ligand="(S)-6-hydroxynicotine"
FT /ligand_id="ChEBI:CHEBI:58182"
FT /evidence="ECO:0000269|PubMed:20006620,
FT ECO:0000269|PubMed:21383134, ECO:0007744|PDB:3K7Q,
FT ECO:0007744|PDB:3NG7, ECO:0007744|PDB:3NGC,
FT ECO:0007744|PDB:3NH3"
FT MUTAGEN 166
FT /note="N->A: 30-fold decrease in kcat/Km and kred for (S)-
FT 6-hydroxynicotine, with larger effects on the kcat/Km value
FT for (S)-6-hydroxynornicotine."
FT /evidence="ECO:0000269|PubMed:28080034"
FT MUTAGEN 187
FT /note="H->N: 3-fold decrease in kcat with (S)-6-
FT hydroxynicotine as substrate."
FT /evidence="ECO:0000269|PubMed:26744768"
FT MUTAGEN 187
FT /note="H->Q: No change in kcat with (S)-6-hydroxynicotine
FT as substrate."
FT /evidence="ECO:0000269|PubMed:26744768"
FT MUTAGEN 287
FT /note="K->M: 10-fold decrease in kcat/Km and kred for (S)-
FT 6-hydroxynicotine. 6000-fold decrease in the kcat/Km value
FT for oxygen."
FT /evidence="ECO:0000269|PubMed:28080034"
FT MUTAGEN 300
FT /note="E->Q: 1.3-fold increase in kcat with (S)-6-
FT hydroxynicotine as substrate."
FT /evidence="ECO:0000269|PubMed:26744768"
FT MUTAGEN 311
FT /note="Y->F: 4-fold decrease in kcat/Km and kred for (S)-6-
FT hydroxynicotine, with larger effects on the kcat/Km value
FT for (S)-6-hydroxynornicotine."
FT /evidence="ECO:0000269|PubMed:28080034"
FT MUTAGEN 407
FT /note="Y->F: No change in kcat with (S)-6-hydroxynicotine
FT as substrate."
FT /evidence="ECO:0000269|PubMed:26744768"
FT CONFLICT 304
FT /note="D -> G (in Ref. 2; CAA11306)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 425 AA; 46335 MW; 7B358BFFD90B1BA2 CRC64;
MYDAIVVGGG FSGLKAARDL TNAGKKVLLL EGGERLGGRA YSRESRNVPG LRVEIGGAYL
HRKHHPRLAA ELDRYGIPTA AASEFTSFRH RLGPTAVDQA FPIPGSEAVA VEAATYTLLR
DAHRIDLEKG LENQDLEDLD IPLNEYVDKL DLPPVSRQFL LAWAWNMLGQ PADQASALWM
LQLVAAHHYS ILGVVLSLDE VFSNGSADLV DAMSQEIPEI RLQTVVTGID QSGDVVNVTV
KDGHAFQAHS VIVATPMNTW RRIVFTPALP ERRRSVIEEG HGGQGLKILI HVRGAEAGIE
CVGDGIFPTL YDYCEVSESE RLLVAFTDSG SFDPTDIGAV KDAVLYYLPE VEVLGIDYHD
WIADPLFEGP WVAPRVGQFS RVHKELGEPA GRIHFVGSDV SLEFPGYIEG ALETAECAVN
AILHS
