RG1_RAUSE
ID RG1_RAUSE Reviewed; 540 AA.
AC Q9SPP9;
DT 11-DEC-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 95.
DE RecName: Full=Raucaffricine-O-beta-D-glucosidase;
DE Short=Raucaffricine beta-glucosidase;
DE Short=RsRG;
DE EC=3.2.1.125;
DE AltName: Full=Vomilenine glucosyltransferase;
DE Short=RsVGT;
DE EC=2.4.1.219;
GN Name=RG; Synonyms=VGT;
OS Rauvolfia serpentina (Serpentine wood) (Ophioxylon serpentinum).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Gentianales; Apocynaceae; Rauvolfioideae; Vinceae;
OC Rauvolfiinae; Rauvolfia.
OX NCBI_TaxID=4060;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 80-111; 319-326; 403-408;
RP 410-425 AND 428-438, FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RC STRAIN=BENTH. ex KURZ; TISSUE=Protoplast;
RX PubMed=10975500; DOI=10.1016/s0031-9422(00)00175-8;
RA Warzecha H., Gerasimenko I., Kutchan T.M., Stoeckigt J.;
RT "Molecular cloning and functional bacterial expression of a plant
RT glucosidase specifically involved in alkaloid biosynthesis.";
RL Phytochemistry 54:657-666(2000).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) OF 1-513 IN COMPLEX WITH SUBSTRATE,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF GLU-186; THR-189;
RP HIS-193; TYR-200; SER-390; TRP-392; GLU-420; GLU-476 AND PHE-485.
RX PubMed=22004291; DOI=10.1021/cb200267w;
RA Xia L., Ruppert M., Wang M., Panjikar S., Lin H., Rajendran C.,
RA Barleben L., Stoeckigt J.;
RT "Structures of alkaloid biosynthetic glucosidases decode substrate
RT specificity.";
RL ACS Chem. Biol. 7:226-234(2012).
CC -!- FUNCTION: Glucosidase specifically involved in alkaloid biosynthesis
CC leading to the accumulation of several alkaloids, including ajmaline,
CC an important plant-derived pharmaceutical used in the treatment of
CC heart disorders. {ECO:0000269|PubMed:10975500}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + raucaffricine = D-glucose + vomilenine;
CC Xref=Rhea:RHEA:14557, ChEBI:CHEBI:4167, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:16408, ChEBI:CHEBI:17400; EC=3.2.1.125;
CC Evidence={ECO:0000269|PubMed:10975500};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=UDP-alpha-D-glucose + vomilenine = H(+) + raucaffricine + UDP;
CC Xref=Rhea:RHEA:19385, ChEBI:CHEBI:15378, ChEBI:CHEBI:16408,
CC ChEBI:CHEBI:17400, ChEBI:CHEBI:58223, ChEBI:CHEBI:58885;
CC EC=2.4.1.219; Evidence={ECO:0000269|PubMed:10975500};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.3 mM for raucaffricine (at pH 5 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:10975500, ECO:0000269|PubMed:22004291};
CC KM=1.8 mM for strictosidine (at pH 5 and 28 degrees Celsius)
CC {ECO:0000269|PubMed:10975500, ECO:0000269|PubMed:22004291};
CC Vmax=0.5 nmol/sec/ug enzyme with raucaffricine as substrate (at pH 5
CC and 28 degrees Celsius) {ECO:0000269|PubMed:10975500,
CC ECO:0000269|PubMed:22004291};
CC Vmax=2.6 pmol/sec/ug enzyme with strictosidine as substrate (at pH 5
CC and 28 degrees Celsius) {ECO:0000269|PubMed:10975500,
CC ECO:0000269|PubMed:22004291};
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 1 family. {ECO:0000305}.
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DR EMBL; AF149311; AAF03675.1; -; mRNA.
DR PDB; 3U57; X-ray; 2.43 A; A/B=1-513.
DR PDB; 3U5U; X-ray; 2.20 A; A/B=1-513.
DR PDB; 3U5Y; X-ray; 2.30 A; A/B=1-513.
DR PDB; 3ZJ6; X-ray; 2.40 A; A/B=1-540.
DR PDB; 4A3Y; X-ray; 2.15 A; A/B=1-540.
DR PDB; 4ATD; X-ray; 2.10 A; A/B=1-513.
DR PDB; 4ATL; X-ray; 2.52 A; A/B=1-513.
DR PDB; 4EK7; X-ray; 2.30 A; A/B=1-513.
DR PDBsum; 3U57; -.
DR PDBsum; 3U5U; -.
DR PDBsum; 3U5Y; -.
DR PDBsum; 3ZJ6; -.
DR PDBsum; 4A3Y; -.
DR PDBsum; 4ATD; -.
DR PDBsum; 4ATL; -.
DR PDBsum; 4EK7; -.
DR AlphaFoldDB; Q9SPP9; -.
DR SMR; Q9SPP9; -.
DR CAZy; GH1; Glycoside Hydrolase Family 1.
DR KEGG; ag:AAF03675; -.
DR BRENDA; 3.2.1.125; 5309.
DR GO; GO:0050247; F:raucaffricine beta-glucosidase activity; IDA:UniProtKB.
DR GO; GO:0050506; F:vomilenine glucosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0009821; P:alkaloid biosynthetic process; IDA:UniProtKB.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR InterPro; IPR001360; Glyco_hydro_1.
DR InterPro; IPR033132; Glyco_hydro_1_N_CS.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR PANTHER; PTHR10353; PTHR10353; 1.
DR Pfam; PF00232; Glyco_hydro_1; 1.
DR PRINTS; PR00131; GLHYDRLASE1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR PROSITE; PS00653; GLYCOSYL_HYDROL_F1_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Direct protein sequencing; Glycosidase;
KW Hydrolase; Transferase.
FT CHAIN 1..540
FT /note="Raucaffricine-O-beta-D-glucosidase"
FT /id="PRO_0000418400"
FT ACT_SITE 186
FT /note="Proton donor"
FT ACT_SITE 420
FT /note="Nucleophile"
FT BINDING 36
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22004291"
FT BINDING 140
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 185..186
FT /ligand="substrate"
FT BINDING 193
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22004291"
FT BINDING 347
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 392
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22004291"
FT BINDING 420
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22004291"
FT BINDING 469
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:22004291"
FT BINDING 476..477
FT /ligand="substrate"
FT SITE 390
FT /note="Directs the conformation of W-392"
FT SITE 392
FT /note="Controls the gate shape and acceptance of
FT substrates"
FT MUTAGEN 186
FT /note="E->D,Q: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 189
FT /note="T->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 193
FT /note="H->A: Reduced activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 200
FT /note="Y->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 390
FT /note="S->G: Reduced activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 392
FT /note="W->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 420
FT /note="E->Q: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 476
FT /note="E->A,L: Loss of activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT MUTAGEN 485
FT /note="F->Y: Reduced activity."
FT /evidence="ECO:0000269|PubMed:22004291"
FT HELIX 14..16
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 19..21
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 27..31
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 34..37
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 50..57
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 59..61
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 63..65
FT /evidence="ECO:0007829|PDB:4ATD"
FT TURN 68..72
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 74..88
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 91..96
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 99..102
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 108..110
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 114..129
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 133..141
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 145..151
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 153..155
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 159..174
FT /evidence="ECO:0007829|PDB:4ATD"
FT TURN 175..177
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 180..185
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 187..195
FT /evidence="ECO:0007829|PDB:4ATD"
FT TURN 235..237
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 238..260
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 262..265
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 268..282
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 286..299
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 301..304
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 306..309
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 314..320
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 321..323
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 329..335
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 340..354
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 367..370
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 373..376
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 378..381
FT /evidence="ECO:0007829|PDB:3U5U"
FT STRAND 382..385
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 398..411
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 414..421
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 433..436
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 440..458
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 463..469
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 477..479
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 482..484
FT /evidence="ECO:0007829|PDB:3ZJ6"
FT STRAND 487..490
FT /evidence="ECO:0007829|PDB:4ATD"
FT TURN 492..496
FT /evidence="ECO:0007829|PDB:4ATD"
FT STRAND 498..500
FT /evidence="ECO:0007829|PDB:4ATD"
FT HELIX 502..511
FT /evidence="ECO:0007829|PDB:4ATD"
SQ SEQUENCE 540 AA; 60934 MW; 543A24654A4D5E7D CRC64;
MATQSSAVID SNDATRISRS DFPADFIMGT GSSAYQIEGG ARDGGRGPSI WDTFTHRRPD
MIRGGTNGDV AVDSYHLYKE DVNILKNLGL DAYRFSISWS RVLPGGRLSG GVNKEGINYY
NNLIDGLLAN GIKPFVTLFH WDVPQALEDE YGGFLSPRIV DDFCEYAELC FWEFGDRVKH
WMTLNEPWTF SVHGYATGLY APGRGRTSPE HVNHPTVQHR CSTVAPQCIC STGNPGTEPY
WVTHHLLLAH AAAVELYKNK FQRGQEGQIG ISHATQWMEP WDENSASDVE AAARALDFML
GWFMEPITSG DYPKSMKKFV GSRLPKFSPE QSKMLKGSYD FVGLNYYTAS YVTNASTNSS
GSNNFSYNTD IHVTYETDRN GVPIGPQSGS DWLLIYPEGI RKILVYTKKT YNVPLIYVTE
NGVDDVKNTN LTLSEARKDS MRLKYLQDHI FNVRQAMNDG VNVKGYFAWS LLDNFEWGEG
YGVRFGIIHI DYNDNFARYP KDSAVWLMNS FHKNISKLPA VKRSIREDDE EQVSSKRLRK
