NEPS1_NEPRA
ID NEPS1_NEPRA Reviewed; 271 AA.
AC A0A3Q8GL18;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 10-APR-2019, sequence version 1.
DT 03-AUG-2022, entry version 12.
DE RecName: Full=(+)-cis,trans-nepetalactol synthase NEPS1 {ECO:0000305};
DE EC=5.5.1.34 {ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
DE AltName: Full=Nepetalactol-related short-chain dehydrogenase {ECO:0000303|PubMed:30531909};
DE Short=Nepetalactol dehydrogenase {ECO:0000303|PubMed:30531909};
DE EC=1.1.1.419 {ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
DE AltName: Full=Nepetalactol-related short-chain reductase 1 {ECO:0000303|PubMed:30531909};
DE Short=Nepetalactol-related SDR1 {ECO:0000303|PubMed:30531909};
DE Short=NmNEPS1 {ECO:0000303|PubMed:30531909};
GN Name=NEPS1 {ECO:0000303|PubMed:30531909};
OS Nepeta racemosa (Catmint) (Raceme catnip).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Lamiales; Lamiaceae; Nepetoideae; Mentheae; Nepetinae;
OC Nepeta.
OX NCBI_TaxID=54731;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF
RP ASN-125; THR-152; THR-153; THR-154; PRO-155; LEU-156; TYR-167; LYS-171;
RP SER-198; VAL-199 AND THR-202, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=30531909; DOI=10.1038/s41589-018-0185-2;
RA Lichman B.R., Kamileen M.O., Titchiner G.R., Saalbach G., Stevenson C.E.M.,
RA Lawson D.M., O'Connor S.E.;
RT "Uncoupled activation and cyclization in catmint reductive terpenoid
RT biosynthesis.";
RL Nat. Chem. Biol. 15:71-79(2019).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=30664302; DOI=10.1002/chem.201805412;
RA Lichman B.R., O'Connor S.E., Kries H.;
RT "Biocatalytic strategies towards [4+2] cycloadditions.";
RL Chemistry 25:6864-6877(2019).
CC -!- FUNCTION: Bifunctional enzyme that possesses cyclase and dehydrogenase
CC activities (PubMed:30531909, PubMed:30664302). Functions as a non-
CC oxidoreductive cyclase to promote the formation of cis-trans-
CC nepetalactol (PubMed:30531909, PubMed:30664302). Functions as
CC dehydrogenase to oxidize cis-cis-nepetalactol and cis-trans-
CC nepetalactol into nepetalactones, metabolites that are both insect-
CC repellent and have euphoric effect in cats (PubMed:30531909,
CC PubMed:30664302). Binds NAD(+) as classical short-chain
CC dehydrogenase/reductase (SDR), but does not utilize it for its redox-
CC neutral cyclase activity (By similarity).
CC {ECO:0000250|UniProtKB:A0A3Q8GLE8, ECO:0000269|PubMed:30531909,
CC ECO:0000269|PubMed:30664302}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-8-oxocitronellyl enol = cis-trans-nepetalactol;
CC Xref=Rhea:RHEA:61416, ChEBI:CHEBI:71494, ChEBI:CHEBI:144481;
CC EC=5.5.1.34; Evidence={ECO:0000269|PubMed:30531909,
CC ECO:0000269|PubMed:30664302};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:61417;
CC Evidence={ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cis-cis-nepetalactol + NAD(+) = cis-cis-nepetalactone + H(+) +
CC NADH; Xref=Rhea:RHEA:61424, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:144482, ChEBI:CHEBI:144485;
CC EC=1.1.1.419; Evidence={ECO:0000269|PubMed:30531909,
CC ECO:0000269|PubMed:30664302};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:61425;
CC Evidence={ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cis-trans-nepetalactol + NAD(+) = cis-trans-nepetalactone +
CC H(+) + NADH; Xref=Rhea:RHEA:61428, ChEBI:CHEBI:7518,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:71494; EC=1.1.1.419;
CC Evidence={ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:61429;
CC Evidence={ECO:0000269|PubMed:30531909, ECO:0000269|PubMed:30664302};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.6 uM for cis-trans-nepetalactol (in the presence of NAD(+) at pH
CC 8.0 and 25 degrees Celsius) {ECO:0000269|PubMed:30531909};
CC KM=4.9 uM for cis-cis-nepetalactol (in the presence of NAD(+) at pH
CC 8.0 and 25 degrees Celsius) {ECO:0000269|PubMed:30531909};
CC KM=3.5 uM for NAD(+) (in the presence of cis-trans-nepetalactol at pH
CC 8.0 and 25 degrees Celsius) {ECO:0000269|PubMed:30531909};
CC Note=kcat is 0.148 sec(-1) with cis-trans-nepetalactol as substrate
CC (in the presence of NAD(+) at pH 8.0 and 25 degrees Celsius). kcat is
CC 0.32 sec(-1) with cis-cis-nepetalactol as substrate (in the presence
CC of NAD(+) at pH 8.0 and 25 degrees Celsius). kcat is 0.173 sec(-1)
CC with NAD(+) as substrate (in the presence of cis-trans-nepetalactol
CC at pH 8.0 and 25 degrees Celsius).;
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
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DR EMBL; MG677124; AXF35971.1; -; mRNA.
DR AlphaFoldDB; A0A3Q8GL18; -.
DR SMR; A0A3Q8GL18; -.
DR KEGG; ag:AXF35971; -.
DR BRENDA; 1.1.1.419; 12895.
DR BRENDA; 5.5.1.34; 12895.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR002347; SDR_fam.
DR PRINTS; PR00081; GDHRDH.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW Isomerase; NAD; Oxidoreductase.
FT CHAIN 1..271
FT /note="(+)-cis,trans-nepetalactol synthase NEPS1"
FT /id="PRO_0000449299"
FT ACT_SITE 167
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:30531909"
FT BINDING 24..30
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A3Q8GLE8"
FT BINDING 49..51
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A3Q8GLE8"
FT BINDING 72..73
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A3Q8GLE8"
FT BINDING 99
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A3Q8GLE8"
FT BINDING 154
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:30531909"
FT BINDING 167
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000305|PubMed:30531909"
FT BINDING 167
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:30531909"
FT BINDING 171
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000305|PubMed:30531909"
FT BINDING 200..205
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000305|PubMed:30531909"
FT MUTAGEN 125
FT /note="N->A: Strongly reduced cis-trans-nepetalactone
FT levels."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 152
FT /note="T->N: Absence of cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 153
FT /note="T->A: Almost normal cis-trans-nepetalactone levels."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 154
FT /note="T->G: Loss of dehydrogenase activity and strongly
FT enhanced cis-trans-nepetalactol levels associated with a
FT huge increase in Km for cis-trans-nepetalactol."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 155
FT /note="P->S: Strongly reduced cis-trans-nepetalactone
FT levels."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 156
FT /note="L->S: Reduced dehydrogenase activity and absence of
FT cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 167
FT /note="Y->F: Absence of cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 171
FT /note="K->M: Absence of cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 198
FT /note="S->M: Absence of cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 199
FT /note="V->A: Almost normal cis-trans-nepetalactone levels."
FT /evidence="ECO:0000269|PubMed:30531909"
FT MUTAGEN 202
FT /note="T->A: Absence of cis-trans-nepetalactone."
FT /evidence="ECO:0000269|PubMed:30531909"
SQ SEQUENCE 271 AA; 28683 MW; 102C4310CC50E98C CRC64;
MASTANPMQV MKKKLEGKVV IVTGGASGIG QTAARVFAQH GARAVVIADI QSEVGKSVAK
SIGDPCCYVQ CDVSDEEEVK SMIEWTASAY GGLDMMFSNV GIMSKSAQTV MDLDLLEFDK
VMRVNARGMA ACLKHAARKM VELGTRGTII CTTTPLSSRG GQSMTDYAMS KHAVMGLVRS
ASIQLGAHGI RVNCVTPSVV LTPLAQRMGL ATPDDFHTHF GNFTSLKGVY LTPEQVAEAV
VYLASDDAAF ITGHDLVLDG GLLCLPFFAP S
