NCS_THLFG
ID NCS_THLFG Reviewed; 210 AA.
AC Q67A25;
DT 20-JAN-2009, integrated into UniProtKB/Swiss-Prot.
DT 11-OCT-2004, sequence version 1.
DT 03-AUG-2022, entry version 56.
DE RecName: Full=S-norcoclaurine synthase;
DE EC=4.2.1.78 {ECO:0000269|PubMed:15447655, ECO:0000269|PubMed:17696451, ECO:0000269|PubMed:19004827};
DE Flags: Precursor;
OS Thalictrum flavum subsp. glaucum (Yellow meadow rue).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Ranunculales; Ranunculaceae; Thalictroideae;
OC Thalictrum.
OX NCBI_TaxID=150095;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 112-120 AND 157-169, TISSUE
RP SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, AND CATALYTIC ACTIVITY.
RX PubMed=15447655; DOI=10.1111/j.1365-313x.2004.02210.x;
RA Samanani N., Liscombe D.K., Facchini P.J.;
RT "Molecular cloning and characterization of norcoclaurine synthase, an
RT enzyme catalyzing the first committed step in benzylisoquinoline alkaloid
RT biosynthesis.";
RL Plant J. 40:302-313(2004).
RN [2]
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, AND CATALYTIC ACTIVITY.
RX PubMed=17696451; DOI=10.1021/bi700752n;
RA Luk L.Y.P., Bunn S., Liscombe D.K., Facchini P.J., Tanner M.E.;
RT "Mechanistic studies on norcoclaurine synthase of benzylisoquinoline
RT alkaloid biosynthesis: an enzymatic Pictet-Spengler reaction.";
RL Biochemistry 46:10153-10161(2007).
RN [3]
RP PROTEIN SEQUENCE OF 30-34, MASS SPECTROMETRY OF 30-210, AND FUNCTION.
RX PubMed=17900926; DOI=10.1016/j.pep.2007.07.010;
RA Berkner H., Engelhorn J., Liscombe D.K., Schweimer K., Woehrl B.M.,
RA Facchini P.J., Roesch P., Matecko I.;
RT "High-yield expression and purification of isotopically labeled
RT norcoclaurine synthase, a Bet v 1-homologous enzyme, from Thalictrum flavum
RT for NMR studies.";
RL Protein Expr. Purif. 56:197-204(2007).
RN [4]
RP FUNCTION, SUBUNIT, AND 3D-STRUCTURE MODELING OF 30-210.
RX PubMed=18384289; DOI=10.1042/bj20080306;
RA Berkner H., Schweimer K., Matecko I., Rosch P.;
RT "Conformation, catalytic site, and enzymatic mechanism of the PR10
RT allergen-related enzyme norcoclaurine synthase.";
RL Biochem. J. 413:281-290(2008).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS), AND CRYSTALLIZATION.
RX PubMed=18391427; DOI=10.1107/s1744309108005678;
RA Pasquo A., Bonamore A., Franceschini S., Macone A., Boffi A., Ilari A.;
RT "Cloning, expression, crystallization and preliminary X-ray data analysis
RT of norcoclaurine synthase from Thalictrum flavum.";
RL Acta Crystallogr. F 64:281-283(2008).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.09 ANGSTROMS) OF 17-210 APOPROTEIN AND IN COMPLEX
RP WITH DOPAMINE AND SUBSTRATE ANALOG P-HYDROXYBENZALDEHYDE,
RP BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF TYR-108; GLU-110 AND LYS-122,
RP AND CATALYTIC ACTIVITY.
RX PubMed=19004827; DOI=10.1074/jbc.m803738200;
RA Ilari A., Franceschini S., Bonamore A., Arenghi F., Botta B., Macone A.,
RA Pasquo A., Bellucci L., Boffi A.;
RT "Structural basis of enzymatic (S)-norcoclaurine biosynthesis.";
RL J. Biol. Chem. 284:897-904(2009).
CC -!- FUNCTION: Involved in the biosynthesis of the common precursor of all
CC benzylisoquinoline alkaloids such as morphine, sanguinarine, codeine or
CC berberine. Condenses dopamine and 4-hydroxyphenylacetaldehyde.
CC {ECO:0000269|PubMed:17696451, ECO:0000269|PubMed:17900926,
CC ECO:0000269|PubMed:18384289}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(4-hydroxyphenyl)acetaldehyde + dopamine = (S)-norcoclaurine +
CC H2O; Xref=Rhea:RHEA:16173, ChEBI:CHEBI:15377, ChEBI:CHEBI:15621,
CC ChEBI:CHEBI:58253, ChEBI:CHEBI:59905; EC=4.2.1.78;
CC Evidence={ECO:0000269|PubMed:15447655, ECO:0000269|PubMed:17696451,
CC ECO:0000269|PubMed:19004827};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=330 uM for 4-hydroxyphenylacetaldehyde
CC {ECO:0000269|PubMed:15447655, ECO:0000269|PubMed:17696451,
CC ECO:0000269|PubMed:19004827};
CC KM=380 uM for dopamine {ECO:0000269|PubMed:15447655,
CC ECO:0000269|PubMed:17696451, ECO:0000269|PubMed:19004827};
CC pH dependence:
CC Optimum pH is 7.0. {ECO:0000269|PubMed:15447655,
CC ECO:0000269|PubMed:17696451, ECO:0000269|PubMed:19004827};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:15447655, ECO:0000269|PubMed:17696451,
CC ECO:0000269|PubMed:19004827};
CC -!- SUBUNIT: Concentration-dependent dimerization, but mainly monomeric at
CC concentrations around 10 uM. {ECO:0000269|PubMed:18384289,
CC ECO:0000269|PubMed:19004827}.
CC -!- TISSUE SPECIFICITY: Expressed most abundantly in the rhizomes and to a
CC lesser extent in petioles, roots, leaves and flower buds.
CC {ECO:0000269|PubMed:15447655}.
CC -!- MASS SPECTROMETRY: Mass=21179.5; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:17900926};
CC -!- MISCELLANEOUS: May be a member of a small family of three to five
CC members.
CC -!- SIMILARITY: Belongs to the BetVI family. {ECO:0000305}.
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DR EMBL; AY376412; AAR22502.1; -; mRNA.
DR PDB; 2VNE; X-ray; 2.72 A; A/B=17-210.
DR PDB; 2VQ5; X-ray; 2.09 A; A/B=17-210.
DR PDB; 5N8Q; X-ray; 2.00 A; A/B/C=34-196.
DR PDB; 5NON; X-ray; 1.85 A; A/B/C=34-196.
DR PDB; 6RP3; X-ray; 1.81 A; A=34-196.
DR PDB; 6Z82; X-ray; 2.30 A; A=34-196.
DR PDBsum; 2VNE; -.
DR PDBsum; 2VQ5; -.
DR PDBsum; 5N8Q; -.
DR PDBsum; 5NON; -.
DR PDBsum; 6RP3; -.
DR PDBsum; 6Z82; -.
DR AlphaFoldDB; Q67A25; -.
DR SMR; Q67A25; -.
DR KEGG; ag:AAR22502; -.
DR BRENDA; 4.2.1.78; 6259.
DR EvolutionaryTrace; Q67A25; -.
DR GO; GO:0050474; F:(S)-norcoclaurine synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0006952; P:defense response; IEA:InterPro.
DR Gene3D; 3.30.530.20; -; 1.
DR InterPro; IPR000916; Bet_v_I/MLP.
DR InterPro; IPR023393; START-like_dom_sf.
DR Pfam; PF00407; Bet_v_1; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Direct protein sequencing; Lyase;
KW Signal.
FT SIGNAL 1..19
FT /evidence="ECO:0000255"
FT CHAIN 20..210
FT /note="S-norcoclaurine synthase"
FT /id="PRO_0000358941"
FT ACT_SITE 122
FT /note="Proton donor"
FT /evidence="ECO:0000269|PubMed:19004827,
FT ECO:0007744|PDB:2VQ5"
FT BINDING 108..110
FT /ligand="dopamine"
FT /ligand_id="ChEBI:CHEBI:59905"
FT /evidence="ECO:0000269|PubMed:19004827,
FT ECO:0007744|PDB:2VQ5"
FT BINDING 141
FT /ligand="(4-hydroxyphenyl)acetaldehyde"
FT /ligand_id="ChEBI:CHEBI:15621"
FT /evidence="ECO:0000269|PubMed:19004827,
FT ECO:0007744|PDB:2VQ5"
FT MUTAGEN 108
FT /note="Y->F: Partial loss of activity."
FT /evidence="ECO:0000269|PubMed:19004827"
FT MUTAGEN 110
FT /note="E->A: Partial loss of activity."
FT /evidence="ECO:0000269|PubMed:19004827"
FT MUTAGEN 122
FT /note="K->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:19004827"
FT STRAND 21..24
FT /evidence="ECO:0007829|PDB:2VQ5"
FT HELIX 37..39
FT /evidence="ECO:0007829|PDB:2VQ5"
FT STRAND 42..54
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 56..63
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 68..75
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 76..78
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 80..90
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 94..99
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 104..116
FT /evidence="ECO:0007829|PDB:6RP3"
FT TURN 117..120
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 121..129
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 130..133
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 136..149
FT /evidence="ECO:0007829|PDB:6RP3"
FT STRAND 152..163
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 165..167
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 168..171
FT /evidence="ECO:0007829|PDB:6RP3"
FT TURN 172..174
FT /evidence="ECO:0007829|PDB:6RP3"
FT HELIX 178..194
FT /evidence="ECO:0007829|PDB:6RP3"
SQ SEQUENCE 210 AA; 23341 MW; 87A47AA31FC96325 CRC64;
MMKMEVVFVF LMLLGTINCQ KLILTGRPFL HHQGIINQVS TVTKVIHHEL EVAASADDIW
TVYSWPGLAK HLPDLLPGAF EKLEIIGDGG VGTILDMTFV PGEFPHEYKE KFILVDNEHR
LKKVQMIEGG YLDLGVTYYM DTIHVVPTGK DSCVIKSSTE YHVKPEFVKI VEPLITTGPL
AAMADAISKL VLEHKSKSNS DEIEAAIITV
