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Database: UniProt
Entry: VINSY_RAUSE
LinkDB: VINSY_RAUSE
Original site: 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   22-FEB-2023, entry version 59.
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; Chloramphenicol acetyltransferase-like domain; 2.
DR   InterPro; IPR023213; CAT-like_dom_sf.
DR   PANTHER; PTHR31623; F21J9.9; 1.
DR   PANTHER; PTHR31623:SF100; VINORINE SYNTHASE-LIKE; 1.
DR   Pfam; PF02458; Transferase; 1.
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
//
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