TCS1_CAMSI
ID TCS1_CAMSI Reviewed; 369 AA.
AC Q9FZN8; A5HF92; K9MNV5;
DT 03-MAY-2011, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 53.
DE RecName: Full=3,7-dimethylxanthine N-methyltransferase TCS1 {ECO:0000305};
DE Short=TCS1a {ECO:0000303|PubMed:26773541, ECO:0000303|PubMed:30303011};
DE EC=2.1.1.160 {ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668, ECO:0000269|PubMed:26773541, ECO:0000269|PubMed:30303011};
DE AltName: Full=Caffeine synthase 1 {ECO:0000303|PubMed:10984041};
GN Name=TCS1 {ECO:0000303|PubMed:10984041};
GN Synonyms=TCS1A {ECO:0000303|PubMed:26773541, ECO:0000303|PubMed:30303011};
OS Camellia sinensis (Tea).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; Ericales; Theaceae; Camellia.
OX NCBI_TaxID=4442;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=10984041; DOI=10.1038/35023072;
RA Kato M., Mizuno K., Crozier A., Fujimura T., Ashihara H.;
RT "Caffeine synthase gene from tea leaves.";
RL Nature 406:956-957(2000).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC TISSUE=Leaf;
RA Deng W.W., Jiang C.J., Wang Z.X., Lee Y.Y., Shi R.J., Zhu L., Ye A.H.,
RA Yu M.;
RT "Cloning and analysis of Camellia sinensis TCS genomic DNA sequence for
RT caffeine synthase.";
RL Submitted (MAR-2007) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Jin J.Q., Chen L., Ma C.L., Yao M.Z.;
RT "Cloning and analysis of Camellia sinensis TCS genomic DNA sequence for
RT caffeine synthase.";
RL Submitted (AUG-2012) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP PROTEIN SEQUENCE OF 14-33, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=10364410; DOI=10.1104/pp.120.2.579;
RA Kato M., Mizuno K., Fujimura T., Iwama M., Irie M., Crozier A.,
RA Ashihara H.;
RT "Purification and characterization of caffeine synthase from tea leaves.";
RL Plant Physiol. 120:579-586(1999).
RN [5]
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=12527364; DOI=10.1016/s0014-5793(02)03781-x;
RA Mizuno K., Okuda A., Kato M., Yoneyama N., Tanaka H., Ashihara H.,
RA Fujimura T.;
RT "Isolation of a new dual-functional caffeine synthase gene encoding an
RT enzyme for the conversion of 7-methylxanthine to caffeine from coffee
RT (Coffea arabica L.).";
RL FEBS Lett. 534:75-81(2003).
RN [6]
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=12860386; DOI=10.1016/s0014-5793(03)00670-7;
RA Mizuno K., Kato M., Irino F., Yoneyama N., Fujimura T., Ashihara H.;
RT "The first committed step reaction of caffeine biosynthesis: 7-
RT methylxanthosine synthase is closely homologous to caffeine synthases in
RT coffee (Coffea arabica L.).";
RL FEBS Lett. 547:56-60(2003).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=16333668; DOI=10.1007/s00438-005-0070-z;
RA Yoneyama N., Morimoto H., Ye C.-X., Ashihara H., Mizuno K., Kato M.;
RT "Substrate specificity of N-methyltransferase involved in purine alkaloids
RT synthesis is dependent upon one amino acid residue of the enzyme.";
RL Mol. Genet. Genomics 275:125-135(2006).
RN [8]
RP REVIEW ON CAFFEINE BIOSYNTHESIS.
RX PubMed=18068204; DOI=10.1016/j.phytochem.2007.10.029;
RA Ashihara H., Sano H., Crozier A.;
RT "Caffeine and related purine alkaloids: biosynthesis, catabolism, function
RT and genetic engineering.";
RL Phytochemistry 69:841-856(2008).
RN [9]
RP TISSUE SPECIFICITY, DEVELOPMENTAL STAGE, AND INDUCTION BY DROUGHT.
RX PubMed=20469645; DOI=10.1515/znc-2010-3-413;
RA Kato M., Kitao N., Ishida M., Morimoto H., Irino F., Mizuno K.;
RT "Expression for caffeine biosynthesis and related enzymes in Camellia
RT sinensis.";
RL Z. Naturforsch. C Biosci. 65:245-256(2010).
RN [10]
RP FUNCTION, MUTAGENESIS OF ARG-225; PHE-271; ALA-272 AND VAL-317, AND
RP BIOTECHNOLOGY.
RX PubMed=25133732; DOI=10.1371/journal.pone.0105368;
RA Jin L., Bhuiya M.W., Li M., Liu X., Han J., Deng W., Wang M., Yu O.,
RA Zhang Z.;
RT "Metabolic engineering of Saccharomyces cerevisiae for caffeine and
RT theobromine production.";
RL PLoS ONE 9:e105368-e105368(2014).
RN [11]
RP FUNCTION, MUTAGENESIS OF SER-269, CATALYTIC ACTIVITY, GENE FAMILY, AND
RP NOMENCLATURE.
RX PubMed=26773541; DOI=10.1016/j.plaphy.2015.12.020;
RA Jin J.-Q., Yao M.-Z., Ma C.-L., Ma J.-Q., Chen L.;
RT "Natural allelic variations of TCS1 play a crucial role in caffeine
RT biosynthesis of tea plant and its related species.";
RL Plant Physiol. Biochem. 100:18-26(2016).
RN [12]
RP MUTAGENESIS OF HIS-153 AND PRO-332.
RX PubMed=27116373; DOI=10.1016/j.plaphy.2016.04.032;
RA Jin J.-Q., Yao M.-Z., Ma C.-L., Ma J.-Q., Chen L.;
RT "Association mapping of caffeine content with TCS1 in tea plant and its
RT related species.";
RL Plant Physiol. Biochem. 105:251-259(2016).
RN [13]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=30303011; DOI=10.1021/acs.jafc.8b03433;
RA Jin J.Q., Chai Y.F., Liu Y.F., Zhang J., Yao M.Z., Chen L.;
RT "Hongyacha, a naturally caffeine-free tea plant from Fujian, China.";
RL J. Agric. Food Chem. 66:11311-11319(2018).
CC -!- FUNCTION: Involved in the biosynthesis of caffeine (PubMed:10984041,
CC PubMed:16333668, PubMed:26773541, PubMed:30303011, PubMed:25133732).
CC Catalyzes the conversion of 7-methylxanthine (7mX) to theobromine and
CC of theobromine to caffeine (PubMed:10984041, PubMed:16333668,
CC PubMed:26773541, PubMed:30303011, PubMed:25133732). Has 3-N- and 1-N-
CC methylation activity (PubMed:10984041, PubMed:16333668).
CC {ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668,
CC ECO:0000269|PubMed:25133732, ECO:0000269|PubMed:26773541,
CC ECO:0000269|PubMed:30303011}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7-methylxanthine + S-adenosyl-L-methionine = H(+) + S-
CC adenosyl-L-homocysteine + theobromine; Xref=Rhea:RHEA:24604,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:28946, ChEBI:CHEBI:48991,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789; EC=2.1.1.160;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668,
CC ECO:0000269|PubMed:26773541, ECO:0000269|PubMed:30303011};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:24605;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668,
CC ECO:0000269|PubMed:26773541};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=S-adenosyl-L-methionine + theobromine = caffeine + H(+) + S-
CC adenosyl-L-homocysteine; Xref=Rhea:RHEA:20944, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:27732, ChEBI:CHEBI:28946, ChEBI:CHEBI:57856,
CC ChEBI:CHEBI:59789; EC=2.1.1.160;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668,
CC ECO:0000269|PubMed:26773541, ECO:0000269|PubMed:30303011};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:20945;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668,
CC ECO:0000269|PubMed:26773541};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=1,7-dimethylxanthine + S-adenosyl-L-methionine = caffeine +
CC H(+) + S-adenosyl-L-homocysteine; Xref=Rhea:RHEA:10280,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:25858, ChEBI:CHEBI:27732,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789; EC=2.1.1.160;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:10281;
CC Evidence={ECO:0000269|PubMed:10984041, ECO:0000269|PubMed:16333668};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q9FLN8};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000250|UniProtKB:Q9FLN8};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=24 uM for paraxanthine {ECO:0000269|PubMed:10364410,
CC ECO:0000269|PubMed:12527364, ECO:0000269|PubMed:12860386};
CC KM=186 uM for 7-methylxanthine {ECO:0000269|PubMed:10364410,
CC ECO:0000269|PubMed:12527364, ECO:0000269|PubMed:12860386};
CC KM=344 uM for theobromine {ECO:0000269|PubMed:10364410,
CC ECO:0000269|PubMed:12527364, ECO:0000269|PubMed:12860386};
CC KM=21 uM for S-adenosyl-L-methionine {ECO:0000269|PubMed:10364410,
CC ECO:0000269|PubMed:12527364, ECO:0000269|PubMed:12860386};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|PubMed:10364410,
CC ECO:0000269|PubMed:12527364, ECO:0000269|PubMed:12860386};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:10984041}.
CC -!- TISSUE SPECIFICITY: Expressed in young leaves and flowers.
CC {ECO:0000269|PubMed:20469645}.
CC -!- DEVELOPMENTAL STAGE: Expression declines during leaf development but
CC remains constant throughout the flower development.
CC {ECO:0000269|PubMed:20469645}.
CC -!- INDUCTION: Down-regulated by drought. {ECO:0000269|PubMed:20469645}.
CC -!- BIOTECHNOLOGY: Saccharomyces cerevisiae (Yeast) expressing Coffea
CC arabica (coffee) xanthosine methyltransferase (CaXMT1) and Camellia
CC sinensis (tea) caffeine synthase (TCS1) accumulates caffeine.
CC {ECO:0000269|PubMed:25133732}.
CC -!- SIMILARITY: Belongs to the methyltransferase superfamily. Type-7
CC methyltransferase family. {ECO:0000305}.
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DR EMBL; AB031280; BAB12278.1; -; mRNA.
DR EMBL; EF526217; ABP98983.1; -; Genomic_DNA.
DR EMBL; JX647690; AFV99128.1; -; Genomic_DNA.
DR AlphaFoldDB; Q9FZN8; -.
DR SMR; Q9FZN8; -.
DR KEGG; ag:BAB12278; -.
DR BRENDA; 2.1.1.159; 1084.
DR BRENDA; 2.1.1.160; 1084.
DR SABIO-RK; Q9FZN8; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0102741; F:paraxanthine:S-adenosyl-L-methionine 3-N-methyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0102740; F:theobromine:S-adenosyl-L-methionine 1-N-methyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1200.270; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR005299; MeTrfase_7.
DR InterPro; IPR042086; MeTrfase_capping.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR PANTHER; PTHR31009; PTHR31009; 1.
DR Pfam; PF03492; Methyltransf_7; 1.
DR SUPFAM; SSF53335; SSF53335; 1.
PE 1: Evidence at protein level;
KW Alkaloid metabolism; Direct protein sequencing; Magnesium; Metal-binding;
KW Methyltransferase; S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..369
FT /note="3,7-dimethylxanthine N-methyltransferase TCS1"
FT /id="PRO_0000408310"
FT BINDING 24
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 24
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE70"
FT BINDING 24
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A4GE70"
FT BINDING 27..31
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
FT BINDING 65..66
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:Q9FLN8"
FT BINDING 65
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
FT BINDING 65
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE70"
FT BINDING 66
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 71
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 101..104
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 138..140
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 155..157
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
FT BINDING 156..160
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A0A6C0WW36"
FT BINDING 156..160
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
FT BINDING 177
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9FLN8"
FT BINDING 263
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9FLN8"
FT BINDING 265
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9FLN8"
FT BINDING 266
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q9FLN8"
FT SITE 153
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000269|PubMed:27116373"
FT SITE 225
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000269|PubMed:25133732"
FT SITE 269
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000269|PubMed:26773541"
FT SITE 317
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000269|PubMed:25133732"
FT SITE 332
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000269|PubMed:27116373"
FT MUTAGEN 153
FT /note="H->Y: Increased theobromine synthase and caffeine
FT synthase activities."
FT /evidence="ECO:0000269|PubMed:27116373"
FT MUTAGEN 225
FT /note="R->H: Reduced theobromine accumulation."
FT /evidence="ECO:0000269|PubMed:25133732"
FT MUTAGEN 269
FT /note="S->C: Increased caffeine synthase activity, but
FT reduced theobromine synthase activity."
FT /evidence="ECO:0000269|PubMed:26773541"
FT MUTAGEN 271
FT /note="F->W: Enhanced theobromine synthase and caffeine
FT synthase activities."
FT /evidence="ECO:0000269|PubMed:25133732"
FT MUTAGEN 272
FT /note="A->P: Reduced theobromine accumulation."
FT /evidence="ECO:0000269|PubMed:25133732"
FT MUTAGEN 317
FT /note="V->M: Enhanced theobromine synthase and caffeine
FT synthase activities."
FT /evidence="ECO:0000269|PubMed:25133732"
FT MUTAGEN 332
FT /note="P->H: Reduced theobromine synthase activity, but
FT normal caffeine synthase activity."
FT /evidence="ECO:0000269|PubMed:27116373"
FT CONFLICT 20
FT /note="G -> E (in Ref. 4; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 29
FT /note="S -> Q (in Ref. 4; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 33
FT /note="Q -> V (in Ref. 4; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 129
FT /note="P -> S (in Ref. 2; ABP98983 and 3; AFV99128)"
FT /evidence="ECO:0000305"
FT CONFLICT 300
FT /note="V -> L (in Ref. 2; ABP98983 and 3; AFV99128)"
FT /evidence="ECO:0000305"
FT CONFLICT 332
FT /note="P -> H (in Ref. 2; ABP98983 and 3; AFV99128)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 369 AA; 41272 MW; E6D262087E475576 CRC64;
MELATAGKVN EVLFMNRGEG ESSYAQNSSF TQQVASMAQP ALENAVETLF SRDFHLQALN
AADLGCAAGP NTFAVISTIK RMMEKKCREL NCQTLELQVY LNDLFGNDFN TLFKGLSSEV
IGNKCEEVPC YVMGVPGSFH GRLFPRNSLH LVHSSYSVHW LTQAPKGLTS REGLALNKGK
IYISKTSPPV VREAYLSQFH EDFTMFLNAR SQEVVPNGCM VLILRGRQCS DPSDMQSCFT
WELLAMAIAE LVSQGLIDED KLDTFNIPSY FASLEEVKDI VERDGSFTID HIEGFDLDSV
EMQENDKWVR GEKFTKVVRA FTEPIISNQF GPEIMDKLYD KFTHIVVSDL EAKLPKTTSI
ILVLSKIDG
