TCMO_HELTU
ID TCMO_HELTU Reviewed; 505 AA.
AC Q04468;
DT 01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1993, sequence version 1.
DT 03-AUG-2022, entry version 101.
DE RecName: Full=Trans-cinnamate 4-monooxygenase;
DE EC=1.14.14.91 {ECO:0000269|PubMed:8026495};
DE AltName: Full=Cinnamic acid 4-hydroxylase;
DE Short=C4H;
DE Short=CA4H;
DE AltName: Full=Cytochrome P450 73;
DE AltName: Full=Cytochrome P450C4H;
GN Name=CYP73A1 {ECO:0000303|PubMed:12223655};
GN Synonyms=CYP73 {ECO:0000303|PubMed:8097885};
OS Helianthus tuberosus (Jerusalem artichoke) (Helianthus tomentosus).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; campanulids; Asterales; Asteraceae; Asteroideae;
OC Heliantheae alliance; Heliantheae; Helianthus.
OX NCBI_TaxID=4233;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], AND PROTEIN SEQUENCE OF 1-21.
RC STRAIN=cv. Blanc commun; TISSUE=Tuberous root;
RX PubMed=8097885; DOI=10.1073/pnas.90.9.4102;
RA Teutsch H.G., Hasenfratz M., Lesot A., Stoltz C., Garnier J.-M.,
RA Jeltsch J.-M., Durst F., Werck-Reichhart D.;
RT "Isolation and sequence of a cDNA encoding the Jerusalem artichoke
RT cinnamate 4-hydroxylase, a major plant cytochrome P450 involved in the
RT general phenylpropanoid pathway.";
RL Proc. Natl. Acad. Sci. U.S.A. 90:4102-4106(1993).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8026495; DOI=10.1111/j.1432-1033.1994.tb18931.x;
RA Urban P., Werck-Reichhart D., Teutsch H.G., Durst F., Regnier S.,
RA Kazmaier M., Pompon D.;
RT "Characterization of recombinant plant cinnamate 4-hydroxylase produced in
RT yeast. Kinetic and spectral properties of the major plant P450 of the
RT phenylpropanoid pathway.";
RL Eur. J. Biochem. 222:843-850(1994).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=9398253; DOI=10.1021/bi971575k;
RA Schalk M., Batard Y., Seyer A., Nedelkina S., Durst F., Werck-Reichhart D.;
RT "Design of fluorescent substrates and potent inhibitors of CYP73As, P450s
RT that catalyze 4-hydroxylation of cinnamic acid in higher plants.";
RL Biochemistry 36:15253-15261(1997).
RN [4]
RP INDUCTION.
RX PubMed=12223655; DOI=10.1104/pp.113.3.951;
RA Batard Y., Schalk M., Pierrel M.A., Zimmerlin A., Durst F.,
RA Werck-Reichhart D.;
RT "Regulation of the cinnamate 4-hydroxylase (CYP73A1) in Jerusalem artichoke
RT tubers in response to wounding and chemical treatments.";
RL Plant Physiol. 113:951-959(1997).
RN [5]
RP ACTIVITY REGULATION.
RX PubMed=9733540; DOI=10.1104/pp.118.1.209;
RA Schalk M., Cabello-Hurtado F., Pierrel M.A., Atanossova R., Saindrenan P.,
RA Werck-Reichhart D.;
RT "Piperonylic acid, a selective, mechanism-based inactivator of the trans-
RT cinnamate 4-hydroxylase: A new tool to control the flux of metabolites in
RT the phenylpropanoid pathway.";
RL Plant Physiol. 118:209-218(1998).
RN [6]
RP MUTAGENESIS OF ALA-306 AND PRO-448.
RX PubMed=10320335; DOI=10.1021/bi982989w;
RA Schalk M., Nedelkina S., Schoch G., Batard Y., Werck-Reichhart D.;
RT "Role of unusual amino acid residues in the proximal and distal heme
RT regions of a plant P450, CYP73A1.";
RL Biochemistry 38:6093-6103(1999).
RN [7]
RP MUTAGENESIS OF ARG-101; ARG-103; ASN-302; ILE-303; ARG-366; ARG-368;
RP ILE-371 AND LYS-484.
RX PubMed=12950252; DOI=10.1046/j.1432-1033.2003.03739.x;
RA Schoch G.A., Attias R., Le Ret M., Werck-Reichhart D.;
RT "Key substrate recognition residues in the active site of a plant
RT cytochrome P450, CYP73A1. Homology guided site-directed mutagenesis.";
RL Eur. J. Biochem. 270:3684-3695(2003).
RN [8]
RP FUNCTION.
RX PubMed=14576280; DOI=10.1104/pp.103.020305;
RA Schoch G.A., Attias R., Belghazi M., Dansette P.M., Werck-Reichhart D.;
RT "Engineering of a water-soluble plant cytochrome P450, CYP73A1, and NMR-
RT based orientation of natural and alternate substrates in the active site.";
RL Plant Physiol. 133:1198-1208(2003).
CC -!- FUNCTION: Catalyzes the first oxidative step of the phenylpropanoid
CC pathway in higher plants by transforming trans-cinnamate into p-
CC coumarate (PubMed:8026495, PubMed:9398253, PubMed:14576280). The
CC compounds formed by this pathway are essential components for
CC lignification, pollination, and defense against ultraviolet light,
CC predators and pathogens (PubMed:8026495, PubMed:9398253,
CC PubMed:14576280). Can also use 2-naphthoic acid as substrate
CC (PubMed:9398253). {ECO:0000269|PubMed:14576280,
CC ECO:0000269|PubMed:8026495, ECO:0000269|PubMed:9398253}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(E)-cinnamate + O2 + reduced [NADPH--hemoprotein reductase] =
CC (E)-4-coumarate + H(+) + H2O + oxidized [NADPH--hemoprotein
CC reductase]; Xref=Rhea:RHEA:10608, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:12876, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:15669, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210; EC=1.14.14.91;
CC Evidence={ECO:0000269|PubMed:8026495, ECO:0000269|PubMed:9398253};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:10609;
CC Evidence={ECO:0000269|PubMed:8026495, ECO:0000269|PubMed:9398253};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q94IP1};
CC -!- ACTIVITY REGULATION: Inactivated by piperonylic acid.
CC {ECO:0000269|PubMed:9733540}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.2 uM for trans-cinnamate {ECO:0000269|PubMed:9398253};
CC KM=2.7 uM for 2-naphthoic acid {ECO:0000269|PubMed:9398253};
CC -!- PATHWAY: Phenylpropanoid metabolism; trans-4-coumarate biosynthesis;
CC trans-4-coumarate from trans-cinnamate: step 1/1. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- INDUCTION: By light, wounding, infection, exposure to xenobiotics and
CC fungal elicitor (Probable). Induced by magnesium chloride, aminopyrine,
CC phenobarbital and clofibrate (at protein level) (PubMed:12223655).
CC {ECO:0000269|PubMed:12223655, ECO:0000305|PubMed:8097885}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; Z17369; CAA78982.1; -; mRNA.
DR PIR; A47454; A47454.
DR AlphaFoldDB; Q04468; -.
DR SMR; Q04468; -.
DR ChEMBL; CHEMBL2268006; -.
DR PRIDE; Q04468; -.
DR BRENDA; 1.14.14.91; 2600.
DR UniPathway; UPA00825; UER00789.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0016710; F:trans-cinnamate 4-monooxygenase activity; IEA:UniProtKB-EC.
DR GO; GO:0051762; P:sesquiterpene biosynthetic process; IEA:UniProt.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Heme; Iron; Membrane; Metal-binding;
KW Monooxygenase; Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..505
FT /note="Trans-cinnamate 4-monooxygenase"
FT /id="PRO_0000052246"
FT TRANSMEM 3..23
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 213..218
FT /ligand="(E)-cinnamate"
FT /ligand_id="ChEBI:CHEBI:15669"
FT /evidence="ECO:0000250|UniProtKB:Q94IP1"
FT BINDING 306
FT /ligand="(E)-cinnamate"
FT /ligand_id="ChEBI:CHEBI:15669"
FT /evidence="ECO:0000250|UniProtKB:Q94IP1"
FT BINDING 447
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250|UniProtKB:Q94IP1"
FT MUTAGEN 101
FT /note="R->M: Reduces catalytic activity 500-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 103
FT /note="R->E: Reduces catalytic activity 3-fold and
FT cinnamate binding affinity 8-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 103
FT /note="R->M: Reduces catalytic activity 2-fold and
FT cinnamate binding affinity 2.4-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 302
FT /note="N->D: Reduces catalytic activity 10-fold and
FT cinnamate binding affinity 6.3-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 302
FT /note="N->F: Reduces catalytic activity 200-fold and
FT cinnamate binding affinity 2-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 303
FT /note="I->A: Reduces catalytic activity 1.3-fold and
FT increases cinnamate binding affinity 1.8-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 306
FT /note="A->G: Decreases cinnamate binding affinity and
FT reduces catalytic efficiency 3-fold."
FT /evidence="ECO:0000269|PubMed:10320335"
FT MUTAGEN 366
FT /note="R->K: Reduces catalytic activity 1.7-fold and
FT cinnamate binding affinity 1.5-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 366
FT /note="R->M: Reduces catalytic activity 1000-fold and
FT cinnamate binding affinity 15-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 368
FT /note="R->F: Reduces catalytic activity 2-fold and
FT cinnamate binding affinity 1.5-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
FT MUTAGEN 371
FT /note="I->A: Reduces catalytic activity 9-fold and
FT cinnamate binding affinity 3.5-fold."
FT MUTAGEN 371
FT /note="I->F: Reduces catalytic activity 1111-fold and
FT cinnamate binding affinity 15-fold."
FT MUTAGEN 371
FT /note="I->K: Reduces catalytic activity 91-fold and
FT cinnamate binding affinity 1.6-fold."
FT MUTAGEN 448
FT /note="P->F,I: Reduces dramatically catalytic activity."
FT /evidence="ECO:0000269|PubMed:10320335"
FT MUTAGEN 484
FT /note="K->M: Reduces catalytic activity 2-fold and
FT increases cinnamate binding affinity 1.2-fold."
FT /evidence="ECO:0000269|PubMed:12950252"
SQ SEQUENCE 505 AA; 57920 MW; 9BA009F5554BAFD7 CRC64;
MDLLLIEKTL VALFAAIIGA ILISKLRGKK FKLPPGPIPV PIFGNWLQVG DDLNHRNLTD
LAKRFGEILL LRMGQRNLVV VSSPELAKEV LHTQGVEFGS RTRNVVFDIF TGKGQDMVFT
VYGEHWRKMR RIMTVPFFTN KVVQQYRYGW EAEAAAVVDD VKKNPAAATE GIVIRRRLQL
MMYNNMFRIM FDRRFESEDD PLFLKLKALN GERSRLAQSF EYNYGDFIPI LRPFLRNYLK
LCKEVKDKRI QLFKDYFVDE RKKIGSTKKM DNNQLKCAID HILEAKEKGE INEDNVLYIV
ENINVAAIET TLWSIEWGIA ELVNHPEIQA KLRHELDTKL GPGVQITEPD VQNLPYLQAV
VKETLRLRMA IPLLVPHMNL HDAKLGGFDI PAESKILVNA WWLANNPDQW KKPEEFRPER
FLEEEAKVEA NGNDFRYLPF GVGRRSCPGI ILALPILGIT IGRLVQNFEL LPPPGQSKID
TDEKGGQFSL HILKHSTIVA KPRSF
