CKCS_CAMSB
ID CKCS_CAMSB Reviewed; 369 AA.
AC A0A6C0WW38;
DT 10-FEB-2021, integrated into UniProtKB/Swiss-Prot.
DT 17-JUN-2020, sequence version 1.
DT 03-AUG-2022, entry version 8.
DE RecName: Full=3,7-dimethylxanthine N-methyltransferase TCS1 {ECO:0000303|PubMed:32193380};
DE EC=2.1.1.160 {ECO:0000269|PubMed:32193380};
DE AltName: Full=Caffeine synthase {ECO:0000303|PubMed:32193380};
DE Short=CkCS {ECO:0000303|PubMed:32193380};
GN Name=CS {ECO:0000303|PubMed:32193380};
OS Camellia sinensis var. assamica (Assam tea) (Thea assamica).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; Ericales; Theaceae; Camellia.
OX NCBI_TaxID=261999;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, PATHWAY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=cv. Kucha;
RX PubMed=32193380; DOI=10.1038/s41467-020-15324-7;
RA Zhang Y.-H., Li Y.-F., Wang Y., Tan L., Cao Z.-Q., Xie C., Xie G.,
RA Gong H.-B., Sun W.-Y., Ouyang S.-H., Duan W.-J., Lu X., Ding K.,
RA Kurihara H., Hu D., Zhang Z.-M., Abe I., He R.-R.;
RT "Identification and characterization of N9-methyltransferase involved in
RT converting caffeine into non-stimulatory theacrine in tea.";
RL Nat. Commun. 11:1473-1473(2020).
CC -!- FUNCTION: Involved in the biosynthesis of caffeine in cv. Puer
CC (PubMed:32193380). Involved in the biosynthesis of theacrine in cv.
CC Kucha, a caffeine-like xanthine alkaloid with diverse beneficial
CC biological activities including anti-depressive, sedative, and hypnotic
CC activities, improving learning and memory, increasing exercise
CC activity, and preventing nonalcoholic fatty liver disease
CC (PubMed:32193380). Catalyzes the conversion of 7-methylxanthine (7mX)
CC to theobromine and of theobromine to caffeine (PubMed:32193380). Has 3-
CC N- and 1-N-methylation activity (PubMed:32193380).
CC {ECO:0000269|PubMed:32193380}.
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:32193380};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:10281;
CC Evidence={ECO:0000269|PubMed:32193380};
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:32193380};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:20945;
CC Evidence={ECO:0000269|PubMed:32193380};
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:32193380};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:24605;
CC Evidence={ECO:0000269|PubMed:32193380};
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=71.49 uM for 1,3,7-trimethyluric acid
CC {ECO:0000269|PubMed:32193380};
CC Note=kcat is 0.00012 sec(-1) with 1,3,7-trimethyluric acid as
CC substrate. {ECO:0000269|PubMed:32193380};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:32193380}.
CC -!- MISCELLANEOUS: Caffeine is catabolized to produce theacrine in Camellia
CC sinensis var. assamica cv. Kucha, but not in cv. Puer.
CC {ECO:0000305|PubMed:32193380}.
CC -!- SIMILARITY: Belongs to the methyltransferase superfamily. Type-7
CC methyltransferase family. {ECO:0000305}.
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DR EMBL; MN163829; QIC50342.1; -; mRNA.
DR AlphaFoldDB; A0A6C0WW38; -.
DR SMR; A0A6C0WW38; -.
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; Magnesium; Metal-binding; Methyltransferase;
KW S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..369
FT /note="3,7-dimethylxanthine N-methyltransferase TCS1"
FT /id="PRO_0000451801"
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 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:A4GE70"
FT BINDING 101..104
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
FT BINDING 138..140
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:A4GE69"
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="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:0000250|UniProtKB:Q9FZN8"
FT SITE 225
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000250|UniProtKB:Q9FZN8"
FT SITE 269
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000250|UniProtKB:Q9FZN8"
FT SITE 317
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000250|UniProtKB:Q9FZN8"
FT SITE 332
FT /note="Involved in substrate discrimination"
FT /evidence="ECO:0000250|UniProtKB:Q9FZN8"
SQ SEQUENCE 369 AA; 41577 MW; 9A1B80145FC301C3 CRC64;
MELATREKVN EVLFMNRGEG ESSYAQNSSF TQQVASMAQP ALENAVETLF SKDFHLQALN
AADLGCAAGP NTFAVISTIK RMMEKKCREL NCQTLELQVY LNDLFGNDFN TLFKGLSSEV
IGNKCEEVPC YVMGVPGSFH GRLFPRNSLH LVYSSYSVHW LTQAPKGLTS REGLALNKGK
IYISKTSPPV VREAYLSQFH EDFTMFLNYR SQEMVPNGCM VLILRGRQCF DPSDMQSCFT
WELLALAIAE LVSQGLIDED KLDTFNIPSY FASLEEVKDI VERDGSFTID HIEGFDLDSV
EMQENDKWVR GEKFTKVVRA FSEPIISSQF GHEIMDKLYD KFTHIVVSDL EAKLPKTTSI
ILVLSKIVG
