SALR_PAPSO
ID SALR_PAPSO Reviewed; 311 AA.
AC Q071N0;
DT 14-OCT-2015, integrated into UniProtKB/Swiss-Prot.
DT 31-OCT-2006, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=Salutaridine reductase {ECO:0000303|PubMed:16968522};
DE EC=1.1.1.248 {ECO:0000269|PubMed:16968522, ECO:0000269|PubMed:19648114};
GN Name=SALR {ECO:0000303|PubMed:16968522};
OS Papaver somniferum (Opium poppy).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Ranunculales; Papaveraceae; Papaveroideae;
OC Papaver.
OX NCBI_TaxID=3469 {ECO:0000312|EMBL:ABC47654.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=16968522; DOI=10.1111/j.1365-313x.2006.02860.x;
RA Ziegler J., Voigtlander S., Schmidt J., Kramell R., Miersch O., Ammer C.,
RA Gesell A., Kutchan T.M.;
RT "Comparative transcript and alkaloid profiling in Papaver species
RT identifies a short chain dehydrogenase/reductase involved in morphine
RT biosynthesis.";
RL Plant J. 48:177-192(2006).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, 3D-STRUCTURE MODELING,
RP ACTIVE SITE, AND MUTAGENESIS OF PHE-104; VAL-106; ASP-107; SER-181;
RP THR-182; LEU-185; LYS-186; LEU-266; MET-271; ASN-272 AND ILE-275.
RX PubMed=19648114; DOI=10.1074/jbc.m109.030957;
RA Ziegler J., Brandt W., Geissler R., Facchini P.J.;
RT "Removal of substrate inhibition and increase in maximal velocity in the
RT short chain dehydrogenase/reductase salutaridine reductase involved in
RT morphine biosynthesis.";
RL J. Biol. Chem. 284:26758-26767(2009).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.91 ANGSTROMS) IN COMPLEX WITH NADP, DISULFIDE
RP BONDS, AND ACTIVITY REGULATION.
RX PubMed=21169353; DOI=10.1074/jbc.m110.168633;
RA Higashi Y., Kutchan T.M., Smith T.J.;
RT "Atomic structure of salutaridine reductase from the opium poppy (Papaver
RT somniferum).";
RL J. Biol. Chem. 286:6532-6541(2011).
CC -!- FUNCTION: Involved in biosynthesis of morphinan-type benzylisoquinoline
CC alkaloids. Catalyzes specifically the stereospecific conversion of
CC salutaridine to salutaridinol. {ECO:0000269|PubMed:16968522,
CC ECO:0000269|PubMed:19648114}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(7S)-salutaridinol + NADP(+) = H(+) + NADPH + salutaridine;
CC Xref=Rhea:RHEA:10108, ChEBI:CHEBI:15378, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58061, ChEBI:CHEBI:58349, ChEBI:CHEBI:58463;
CC EC=1.1.1.248; Evidence={ECO:0000269|PubMed:16968522,
CC ECO:0000269|PubMed:19648114};
CC -!- ACTIVITY REGULATION: Strong substrate inhibition (PubMed:19648114). Was
CC thought to be due to mutually exclusive productive and non-productive
CC modes of substrate binding in the active site (PubMed:19648114).
CC Alternatively, SALR may undergo significant conformational changes
CC during catalytic turnover (PubMed:21169353).
CC {ECO:0000269|PubMed:19648114, ECO:0000269|PubMed:21169353}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=80 uM for NADPH {ECO:0000269|PubMed:16968522};
CC KM=198 uM for NADP {ECO:0000269|PubMed:16968522};
CC KM=30.9 uM for salutaridine {ECO:0000269|PubMed:16968522};
CC KM=23 uM for salutaridinol {ECO:0000269|PubMed:16968522};
CC Vmax=9.4 pmol/sec/mg enzyme toward NADPH
CC {ECO:0000269|PubMed:16968522};
CC Vmax=11.6 pmol/sec/mg enzyme toward NADP
CC {ECO:0000269|PubMed:16968522};
CC Vmax=5.8 pmol/sec/mg enzyme with salutaridine as substrate
CC {ECO:0000269|PubMed:16968522};
CC Vmax=10.6 pmol/sec/mg enzyme with salutaridinol as substrate
CC {ECO:0000269|PubMed:16968522};
CC pH dependence:
CC Optimum pH is between 5.5 and 6.0. {ECO:0000269|PubMed:16968522};
CC Temperature dependence:
CC Optimum temperature is between 30 and 35 degrees Celsius.
CC {ECO:0000269|PubMed:16968522};
CC -!- PATHWAY: Alkaloid biosynthesis; morphine biosynthesis.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; DQ316261; ABC47654.1; -; mRNA.
DR PDB; 3O26; X-ray; 1.91 A; A=1-311.
DR PDBsum; 3O26; -.
DR AlphaFoldDB; Q071N0; -.
DR SMR; Q071N0; -.
DR KEGG; ag:ABC47654; -.
DR BioCyc; MetaCyc:MON-12300; -.
DR BRENDA; 1.1.1.248; 4515.
DR UniPathway; UPA00852; -.
DR GO; GO:0050661; F:NADP binding; IDA:UniProtKB.
DR GO; GO:0047037; F:salutaridine reductase (NADPH) activity; IDA:UniProtKB.
DR GO; GO:0097295; P:morphine biosynthetic process; IEP:UniProtKB.
DR CDD; cd05324; carb_red_PTCR-like_SDR_c; 1.
DR InterPro; IPR045313; CBR1-like.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR002347; SDR_fam.
DR Pfam; PF00106; adh_short; 1.
DR PRINTS; PR00081; GDHRDH.
DR PRINTS; PR00080; SDRFAMILY.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Disulfide bond; NAD; NADP;
KW Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..311
FT /note="Salutaridine reductase"
FT /id="PRO_0000433978"
FT ACT_SITE 236
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:21169353"
FT BINDING 21..24
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 44
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 70..71
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 98
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 129
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21169353"
FT BINDING 180
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21169353"
FT BINDING 236
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 240
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT BINDING 267..272
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT DISULFID 263..305
FT /evidence="ECO:0000269|PubMed:21169353,
FT ECO:0007744|PDB:3O26"
FT MUTAGEN 104
FT /note="F->A: Weak substrate inhibition, decreased substrate
FT affinity and increased reaction velocity. Loss of substrate
FT inhibition and doubled maximal velocity; when associated
FT with A-275."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 106
FT /note="V->A: Decreased substrate affinity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 107
FT /note="D->A: Decreased substrate affinity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 181
FT /note="S->A: No effect on activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 182
FT /note="T->A: Slightly decreased affinity, but strongly
FT reduced activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 185
FT /note="L->A: Very low activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 185
FT /note="L->S: Increased substrate affinity, but low
FT activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 185
FT /note="L->V: Reduced activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 186
FT /note="K->V: No effect on activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 266
FT /note="L->A: Decreased substrate affinity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 266
FT /note="L->V: No effect on substrate affinity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 271
FT /note="M->A: Abrogate substrate inhibition. Decreased
FT substrate affinity and decreased reductase activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 272
FT /note="N->A: Abrogate substrate inhibition. Decreased
FT substrate affinity and decreased reductase activity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 275
FT /note="I->A: Weak substrate inhibition, decreased substrate
FT affinity and increased reaction velocity. Loss of substrate
FT inhibition and doubled maximal velocity; when associated
FT with A-104."
FT /evidence="ECO:0000269|PubMed:19648114"
FT MUTAGEN 275
FT /note="I->V: No effect on substrate affinity."
FT /evidence="ECO:0000269|PubMed:19648114"
FT STRAND 14..19
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 23..34
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 38..44
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 46..57
FT /evidence="ECO:0007829|PDB:3O26"
FT TURN 58..60
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 63..68
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 75..89
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 94..97
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 108..118
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 124..128
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 132..135
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 142..152
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 154..167
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 170..172
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 174..178
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 185..187
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 191..198
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 200..202
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 205..220
FT /evidence="ECO:0007829|PDB:3O26"
FT TURN 224..228
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 234..253
FT /evidence="ECO:0007829|PDB:3O26"
FT STRAND 257..262
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 270..272
FT /evidence="ECO:0007829|PDB:3O26"
FT HELIX 280..291
FT /evidence="ECO:0007829|PDB:3O26"
SQ SEQUENCE 311 AA; 34050 MW; 60E9B07FAC93353F CRC64;
MPETCPNTVT KRRCAVVTGG NKGIGFEICK QLSSNGIMVV LTCRDVTKGH EAVEKLKNSN
HENVVFHQLD VTDPIATMSS LADFIKTHFG KLDILVNNAG VAGFSVDADR FKAMISDIGE
DSEELVKIYE KPEAQELMSE TYELAEECLK INYNGVKSVT EVLIPLLQLS DSPRIVNVSS
STGSLKYVSN ETALEILGDG DALTEERIDM VVNMLLKDFK ENLIETNGWP SFGAAYTTSK
ACLNAYTRVL ANKIPKFQVN CVCPGLVKTE MNYGIGNYTA EEGAEHVVRI ALFPDDGPSG
FFYDCSELSA F
