VINSY_RAUSE
ID VINSY_RAUSE Reviewed; 421 AA.
AC Q70PR7;
DT 24-JUL-2007, integrated into UniProtKB/Swiss-Prot.
DT 15-FEB-2005, sequence version 2.
DT 03-AUG-2022, entry version 58.
DE RecName: Full=Vinorine synthase {ECO:0000303|PubMed:15110860};
DE EC=2.3.1.160 {ECO:0000269|PubMed:15110860};
GN Name=ACT {ECO:0000303|PubMed:15110860};
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 2-21; 132-143; 181-195;
RP 219-225 AND 245-254, MUTAGENESIS OF SER-16; SER-29; ASP-32; SER-68; CYS-89;
RP CYS-149; HIS-160; ASP-164; SER-243; ASN-293; ASP-360; ASP-362 AND SER-413,
RP FUNCTION, ACTIVE SITES, BIOPHYSICOCHEMICAL PROPERTIES, CATALYTIC ACTIVITY,
RP AND ACTIVITY REGULATION.
RX PubMed=15110860; DOI=10.1016/j.bmc.2004.02.029;
RA Bayer A., Ma X., Stoeckigt J.;
RT "Acetyltransfer in natural product biosynthesis -- functional cloning and
RT molecular analysis of vinorine synthase.";
RL Bioorg. Med. Chem. 12:2787-2795(2004).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS), ACTIVE SITES, AND SUBUNIT.
RX PubMed=15665331; DOI=10.1074/jbc.m414508200;
RA Ma X., Koepke J., Panjikar S., Fritzsch G., Stoeckigt J.;
RT "Crystal structure of vinorine synthase, the first representative of the
RT BAHD superfamily.";
RL J. Biol. Chem. 280:13576-13583(2005).
CC -!- FUNCTION: Acetyltransferase that catalyzes the formation of vinorine, a
CC precursor of the antiarrhythmic monoterpenoid indole alkaloid ajmaline.
CC Acts on gardneral, but not on polyneuridine aldehyde or N-
CC methylgardneral. {ECO:0000269|PubMed:15110860}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=16-epivellosimine + acetyl-CoA = CoA + vinorine;
CC Xref=Rhea:RHEA:24016, ChEBI:CHEBI:16425, ChEBI:CHEBI:16791,
CC ChEBI:CHEBI:57287, ChEBI:CHEBI:57288; EC=2.3.1.160;
CC Evidence={ECO:0000269|PubMed:15110860};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:24017;
CC Evidence={ECO:0000269|PubMed:15110860};
CC -!- ACTIVITY REGULATION: Complete inhibition by 4-(2-aminoethyl)-
CC benzenesulfonyl fluoride (AEBSF), N-tosyl-L-phenylalanine
CC chloromethylketone (TPCK), Hg(2+) and diethyl-pyrocarbonate (DEPC)
CC (PubMed:15110860). 50% inhibition by N-(N-(L-3-trans-carboxirane-2-
CC carbonyl)-L-leucyl)-agmanitine (E-64), N-alpha-p-tosyl-L-lysine
CC chloromethylketone (TLCK) and phenylmethylsulfonyl fluoride (PMSF)
CC (PubMed:15110860). {ECO:0000269|PubMed:15110860}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=57 uM for Acetyl-CoA {ECO:0000269|PubMed:15110860};
CC KM=7.5 uM for gardneral {ECO:0000269|PubMed:15110860};
CC KM=63 uM for CoA {ECO:0000269|PubMed:15110860};
CC KM=10 uM for vinorine {ECO:0000269|PubMed:15110860};
CC Vmax=3.9 umol/min/mg enzyme for the forward reaction
CC {ECO:0000269|PubMed:15110860};
CC Vmax=44.1 umol/min/mg enzyme for the reverse reaction
CC {ECO:0000269|PubMed:15110860};
CC Note=Except vinorine, no other acetylated alkaloids are deacetylated
CC by the reverse reaction. {ECO:0000269|PubMed:15110860};
CC pH dependence:
CC Optimum pH is 7.8. {ECO:0000269|PubMed:15110860};
CC Temperature dependence:
CC Optimum temperature is 35 degrees Celsius.
CC {ECO:0000269|PubMed:15110860};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:15665331}.
CC -!- SIMILARITY: Belongs to the plant acyltransferase family. {ECO:0000305}.
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DR EMBL; AJ556780; CAD89104.2; -; mRNA.
DR PDB; 2BGH; X-ray; 2.60 A; A/B=1-421.
DR PDBsum; 2BGH; -.
DR AlphaFoldDB; Q70PR7; -.
DR SMR; Q70PR7; -.
DR KEGG; ag:CAD89104; -.
DR BRENDA; 2.3.1.160; 5309.
DR EvolutionaryTrace; Q70PR7; -.
DR GO; GO:0050636; F:vinorine synthase activity; IDA:UniProtKB.
DR GO; GO:0009820; P:alkaloid metabolic process; IDA:UniProtKB.
DR Gene3D; 3.30.559.10; -; 2.
DR InterPro; IPR023213; CAT-like_dom_sf.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Alkaloid metabolism;
KW Direct protein sequencing; Transferase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:15110860"
FT CHAIN 2..421
FT /note="Vinorine synthase"
FT /id="PRO_0000295862"
FT ACT_SITE 160
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:15110860,
FT ECO:0000305|PubMed:15665331"
FT ACT_SITE 362
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:15110860,
FT ECO:0000305|PubMed:15665331"
FT MUTAGEN 16
FT /note="S->A: 29% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 29
FT /note="S->A: 75% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 32
FT /note="D->A: 86% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 68
FT /note="S->A: No effect."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 89
FT /note="C->A: No effect."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 149
FT /note="C->A: 90% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 160
FT /note="H->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 164
FT /note="D->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 243
FT /note="S->A: 83% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 293
FT /note="N->A: 32% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 360
FT /note="D->A: No effect."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 362
FT /note="D->A: 65% reduction of activity."
FT /evidence="ECO:0000269|PubMed:15110860"
FT MUTAGEN 413
FT /note="S->A: No effect."
FT /evidence="ECO:0000269|PubMed:15110860"
FT STRAND 5..14
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 30..33
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 37..46
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 56..70
FT /evidence="ECO:0007829|PDB:2BGH"
FT TURN 71..73
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 75..78
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 79..82
FT /evidence="ECO:0007829|PDB:2BGH"
FT TURN 83..85
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 86..88
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 94..103
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 105..109
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 115..121
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 122..124
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 126..132
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 139..146
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 152..159
FT /evidence="ECO:0007829|PDB:2BGH"
FT TURN 160..162
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 165..179
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 192..196
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 212..221
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 223..232
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 244..263
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 270..278
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 279..281
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 282..284
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 295..302
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 310..313
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 314..320
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 328..340
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 344..346
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 347..353
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 358..360
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 363..365
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 369..372
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 381..386
FT /evidence="ECO:0007829|PDB:2BGH"
FT STRAND 390..400
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 401..406
FT /evidence="ECO:0007829|PDB:2BGH"
FT HELIX 409..412
FT /evidence="ECO:0007829|PDB:2BGH"
SQ SEQUENCE 421 AA; 46828 MW; 3C96D9872358CFA4 CRC64;
MAPQMEKVSE ELILPSSPTP QSLKCYKISH LDQLLLTCHI PFILFYPNPL DSNLDPAQTS
QHLKQSLSKV LTHFYPLAGR INVNSSVDCN DSGVPFVEAR VQAQLSQAIQ NVVELEKLDQ
YLPSAAYPGG KIEVNEDVPL AVKISFFECG GTAIGVNLSH KIADVLSLAT FLNAWTATCR
GETEIVLPNF DLAARHFPPV DNTPSPELVP DENVVMKRFV FDKEKIGALR AQASSASEEK
NFSRVQLVVA YIWKHVIDVT RAKYGAKNKF VVVQAVNLRS RMNPPLPHYA MGNIATLLFA
AVDAEWDKDF PDLIGPLRTS LEKTEDDHNH ELLKGMTCLY ELEPQELLSF TSWCRLGFYD
LDFGWGKPLS ACTTTFPKRN AALLMDTRSG DGVEAWLPMA EDEMAMLPVE LLSLVDSDFS
K
