HLTT_LUPAL
ID HLTT_LUPAL Reviewed; 453 AA.
AC Q5H873; A3FK14;
DT 21-SEP-2011, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2005, sequence version 1.
DT 03-AUG-2022, entry version 40.
DE RecName: Full=13-hydroxylupanine O-tigloyltransferase;
DE EC=2.3.1.93;
DE AltName: Full=(-)-13alpha-hydroxymultiflorine/(+)-13alpha-hydroxylupanine O-tigloyltransferase;
DE Short=HMT/HLTase;
DE AltName: Full=Quinolizidine alkaloid O-tigloyltransferase;
GN Name=HMT/HLT;
OS Lupinus albus (White lupine) (Lupinus termis).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade;
OC genistoids sensu lato; core genistoids; Genisteae; Lupinus.
OX NCBI_TaxID=3870;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 2-9; 15-24; 229-233;
RP 360-371; 383-390 AND 435-441, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, AND TISSUE SPECIFICITY.
RC TISSUE=Root;
RX PubMed=15659437; DOI=10.1093/pcp/pci021;
RA Okada T., Hirai M.Y., Suzuki H., Yamazaki M., Saito K.;
RT "Molecular characterization of a novel quinolizidine alkaloid O-
RT tigloyltransferase: cDNA cloning, catalytic activity of recombinant protein
RT and expression analysis in Lupinus plants.";
RL Plant Cell Physiol. 46:233-244(2005).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND TISSUE SPECIFICITY.
RX AGRICOLA=IND43981835; DOI=10.1111/j.1744-7348.2007.00175.x;
RA Chen Y., Lee L.S., Luckett D.J., Henry R., Hill H., Edwards M.;
RT "A quinolizidine alkaloid O-tigloyltransferase gene in wild and
RT domesticated white lupin (Lupinus albus).";
RL Ann. Appl. Biol. 151:357-362(2007).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, SUBUNIT, AND TISSUE SPECIFICITY.
RX PubMed=8195240; DOI=10.1016/s0021-9258(17)40759-9;
RA Suzuki H., Murakoshi I., Saito K.;
RT "A novel O-tigloyltransferase for alkaloid biosynthesis in plants.
RT Purification, characterization, and distribution in Lupinus plants.";
RL J. Biol. Chem. 269:15853-15860(1994).
CC -!- FUNCTION: Acyl-CoA-dependent acyltransferase involved in the synthesis
CC of lupanine alkaloids. Can use both (-)-13alpha-hydroxymultiflorine and
CC (+)-13alpha-hydroxylupanine as substrates. Lower activity with (-)-
CC 3beta, 13alpha-dihydroxylupanine, but no activity with (+)-epilupinine
CC and (-)-lupinine as substrates. Tigloyl-CoA, benzoyl-CoA and, more
CC slowly, acetyl-CoA, propionyl-CoA and 2-butenoyl-CoA can act as acyl
CC donors. {ECO:0000269|PubMed:15659437, ECO:0000269|PubMed:8195240}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E)-2-methylbut-2-enoyl-CoA + 13-hydroxylupanine = 13-(2-
CC methylcrotonoyloxy)lupanine + CoA; Xref=Rhea:RHEA:12360,
CC ChEBI:CHEBI:57287, ChEBI:CHEBI:57337, ChEBI:CHEBI:58446,
CC ChEBI:CHEBI:58460; EC=2.3.1.93;
CC Evidence={ECO:0000269|PubMed:15659437, ECO:0000269|PubMed:8195240};
CC -!- ACTIVITY REGULATION: Inhibited by N-ethylmaleimide, p-
CC chloromercuribenzoic acid and diethylpyrocarbonate (DEPC).
CC {ECO:0000269|PubMed:15659437, ECO:0000269|PubMed:8195240}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=94 uM for 13alpha-hydroxymultiflorine (with tigloyl-CoA as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC KM=112 uM for 13alpha-hydroxylupanine (with tigloyl-CoA as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC KM=98 uM for tigloyl-CoA (with 13alpha-hydroxymultiflorine as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC KM=359 uM for tigloyl-CoA (with 13alpha-hydroxylupanine as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC KM=93 uM for benzoyl-CoA (with 13alpha-hydroxymultiflorine as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC KM=405 uM for benzoyl-CoA (with 13alpha-hydroxylupanine as
CC cosubstrate) {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:15659437,
CC ECO:0000269|PubMed:8195240};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:8195240}.
CC -!- TISSUE SPECIFICITY: Expressed in roots and hypocotyls. Detected in
CC seeds, leaves and cotyledons, but not in young developing leaves.
CC {ECO:0000269|PubMed:15659437, ECO:0000269|PubMed:8195240,
CC ECO:0000269|Ref.2}.
CC -!- SIMILARITY: Belongs to the plant acyltransferase family. {ECO:0000305}.
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DR EMBL; AB181292; BAD89275.1; -; mRNA.
DR EMBL; EF381744; ABN48480.1; -; Genomic_DNA.
DR AlphaFoldDB; Q5H873; -.
DR SMR; Q5H873; -.
DR KEGG; ag:BAD89275; -.
DR BRENDA; 2.3.1.93; 3089.
DR GO; GO:0047203; F:13-hydroxylupinine O-tigloyltransferase activity; IDA:UniProtKB.
DR GO; GO:0009821; P:alkaloid biosynthetic process; IDA:UniProtKB.
DR Gene3D; 3.30.559.10; -; 2.
DR InterPro; IPR023213; CAT-like_dom_sf.
PE 1: Evidence at protein level;
KW Acyltransferase; Direct protein sequencing; Transferase.
FT CHAIN 1..453
FT /note="13-hydroxylupanine O-tigloyltransferase"
FT /id="PRO_0000412590"
FT ACT_SITE 166
FT /note="Proton acceptor"
FT /evidence="ECO:0000255"
FT ACT_SITE 385
FT /note="Proton acceptor"
FT /evidence="ECO:0000255"
FT CONFLICT 65
FT /note="E -> A (in Ref. 2; ABN48480)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 453 AA; 51181 MW; 5CF8D70509EF12C2 CRC64;
MAPQTQSLVF KVRRNPQELV TPAKPTPKEF KLLSDIDDQT SLRSLTPLVT IYRNNPSMEG
KDPVEIIREA LSKTLVFYYP FAGRLRNGPN GKLMVDCTGE GVIFIEADAD VTLDQFGIDL
HPPFPCFDQL LYDVPGSDGI LDSPLLLIQV TRLKCGGFIF AVRLNHAMCD AIGMSQFMKG
LAEIARGEPK PFILPVWHRE LLCARNPPKV TFIHNEYQKP PHDNNNNNFI LQHSSFFFGP
NELDAIRRLL PYHHSKSTTS DILTAFLWRC RTLALQPENP NHEFRLLYIL NARYGRCSFN
PPLPEGFYGN AFVSPAAIST GEKLCNNPLE YALELMKEAK SKGTEEYVHS VADLMVIKGR
PSYFYNDVGY LEVSDLTKAR FRDVDFGWGK AVYGGATQGY FSSILYVSYT NSKGVEGIMA
LTSLPTKAME RFEKELDDLF KTKDKSQILR SHI
