LOLP1_EPIUN
ID LOLP1_EPIUN Reviewed; 496 AA.
AC Q5MNI1;
DT 23-MAY-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-2005, sequence version 1.
DT 03-AUG-2022, entry version 63.
DE RecName: Full=Cytochrome P450 monooxygenase lolP1 {ECO:0000303|PubMed:15654104};
DE EC=1.-.-.- {ECO:0000305|PubMed:15654104};
DE AltName: Full=Loline biosynthesis cluster 1 protein P {ECO:0000303|PubMed:15654104};
GN Name=lolP1 {ECO:0000303|PubMed:15654104};
GN Synonyms=lolP {ECO:0000303|PubMed:15654104};
OS Epichloe uncinata (Endophyte fungus) (Neotyphodium uncinatum).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Clavicipitaceae; Epichloe.
OX NCBI_TaxID=5050;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], INDUCTION, AND FUNCTION.
RC STRAIN=CBS 102646;
RX PubMed=15654104; DOI=10.1534/genetics.104.035972;
RA Spiering M.J., Moon C.D., Wilkinson H.H., Schardl C.L.;
RT "Gene clusters for insecticidal loline alkaloids in the grass-endophytic
RT fungus Neotyphodium uncinatum.";
RL Genetics 169:1403-1414(2005).
RN [2]
RP FUNCTION.
RX PubMed=15861432; DOI=10.1002/cbic.200400327;
RA Blankenship J.D., Houseknecht J.B., Pal S., Bush L.P., Grossman R.B.,
RA Schardl C.L.;
RT "Biosynthetic precursors of fungal pyrrolizidines, the loline alkaloids.";
RL ChemBioChem 6:1016-1022(2005).
RN [3]
RP FUNCTION.
RX PubMed=16755627; DOI=10.1002/cbic.200600066;
RA Faulkner J.R., Hussaini S.R., Blankenship J.D., Pal S., Branan B.M.,
RA Grossman R.B., Schardl C.L.;
RT "On the sequence of bond formation in loline alkaloid biosynthesis.";
RL ChemBioChem 7:1078-1088(2006).
RN [4]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=CBS 102646;
RX PubMed=18655839; DOI=10.1016/j.fgb.2008.07.001;
RA Spiering M.J., Faulkner J.R., Zhang D.X., Machado C., Grossman R.B.,
RA Schardl C.L.;
RT "Role of the LolP cytochrome P450 monooxygenase in loline alkaloid
RT biosynthesis.";
RL Fungal Genet. Biol. 45:1307-1314(2008).
RN [5]
RP FUNCTION.
RX PubMed=24374065; DOI=10.1016/j.phytochem.2013.11.015;
RA Pan J., Bhardwaj M., Faulkner J.R., Nagabhyru P., Charlton N.D.,
RA Higashi R.M., Miller A.F., Young C.A., Grossman R.B., Schardl C.L.;
RT "Ether bridge formation in loline alkaloid biosynthesis.";
RL Phytochemistry 98:60-68(2014).
RN [6]
RP FUNCTION, AND BIOTECHNOLOGY.
RX PubMed=25531527; DOI=10.1371/journal.pone.0115590;
RA Pan J., Bhardwaj M., Nagabhyru P., Grossman R.B., Schardl C.L.;
RT "Enzymes from fungal and plant origin required for chemical diversification
RT of insecticidal loline alkaloids in grass-Epichloe symbiota.";
RL PLoS ONE 9:E115590-E115590(2014).
RN [7]
RP FUNCTION.
RX PubMed=29537853; DOI=10.1021/acs.biochem.8b00157;
RA Pan J., Bhardwaj M., Zhang B., Chang W.C., Schardl C.L., Krebs C.,
RA Grossman R.B., Bollinger J.M. Jr.;
RT "Installation of the ether bridge of lolines by the iron- and 2-
RT oxoglutarate-dependent oxygenase, lolO: regio- and stereochemistry of
RT sequential hydroxylation and oxacyclization reactions.";
RL Biochemistry 57:2074-2083(2018).
CC -!- FUNCTION: Cytochrome P450 monooxygenase; part of the gene cluster that
CC mediates the biosynthesis of loline alkaloids, potent insecticidal
CC agents composed of a pyrrolizidine ring system and an uncommon ether
CC bridge linking carbons 2 and 7 (PubMed:15654104). Lolines are
CC structurally differentiated by the various modifications of the L-amino
CC group and include norloline, loline, N-methylloline, N-acetylloline, N-
CC acetylnorloline, and N-formylloline (PubMed:15861432, PubMed:25531527).
CC The first committed step is the condensation of O-acetyl-L-homoserine
CC (derived from L-aspartic acid) and L-proline, probably catalyzed by the
CC gamma-type pyridoxal 5'-phosphate(PLP)-dependent enzyme lolC, to give
CC the diamino diacid, NACPP (PubMed:15861432, PubMed:16755627). Ensuing
CC cyclization, decarboxylation, and acetylation steps yield 1-exo-
CC acetamidopyrrolizidine (AcAP)(PubMed:24374065). LolO is required for
CC installation of the ether bridge upon the pathway intermediate, 1-exo-
CC acetamidopyrrolizidine (AcAP) (PubMed:29537853). In sequential 2-
CC oxoglutarate- and O(2)-consuming steps, lolO removes hydrogens from C2
CC and C7 of AcAP to form both carbon-oxygen bonds in N-acetylnorloline
CC (NANL), the precursor to all other lolines (PubMed:24374065,
CC PubMed:29537853). The enzymes lolD, lolE, lolF and lolT have also been
CC proposed to be involved in the ether-bridge installation
CC (PubMed:15654104). Further processing of the exocyclic moiety of NANL
CC by fungal N-acetamidase (LolN), methyltransferase (LolM), and
CC cytochrome P450 (LolP) enzymes, with occasional involvement of a plant
CC acetyltransferase, generates the other known lolines (PubMed:25531527,
CC PubMed:18655839). LolN transforms NANL to norlonine which is
CC monomethylated and dimethylated to respectively lonine and N-
CC methyllonine (NML) by lolM (PubMed:25531527). LolP catalyzes
CC hydroxylation of the methyl group in N-methylloline (NML) and further
CC oxygenation to N-formylloline (NFL) (PubMed:18655839). A plant
CC acetyltransferase is responsible for the acetylation of loline to form
CC N-acetylloline (NAL) (PubMed:25531527). LolA might interact with
CC aspartate kinase to prevent feedback inhibition of its activity by
CC these end products and thereby promote production of L-homoserine from
CC L-aspartate (PubMed:15654104). {ECO:0000269|PubMed:15654104,
CC ECO:0000269|PubMed:15861432, ECO:0000269|PubMed:16755627,
CC ECO:0000269|PubMed:18655839, ECO:0000269|PubMed:24374065,
CC ECO:0000269|PubMed:25531527, ECO:0000269|PubMed:29537853}.
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:P04798};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:18655839}.
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- INDUCTION: Expression is induced in loline alkaloid-producing cultures
CC as well as in planta (PubMed:15654104). {ECO:0000269|PubMed:15654104}.
CC -!- DISRUPTION PHENOTYPE: Leads to elevated levels of N-methylloline (NML)
CC and lacks N-formylloline (NFL)(PubMed:18655839).
CC {ECO:0000269|PubMed:18655839}.
CC -!- BIOTECHNOLOGY: Loline alkaloids show broad-spectrum anti-insect
CC activity, and different lolines may have different biological
CC activities (PubMed:25531527). In vitro tests of NFL, NAL, NML, and
CC semisynthetic loline derivatives with long carbon-chain acylations on
CC the 1-amine have shown that many are effective against both fall
CC armyworm larvae and European corn borer larvae, but the effects seem to
CC differ depending on the modifications (PubMed:25531527). N-Formylloline
CC reduces the weight gain of fall armyworms by deterring feeding, and
CC does not significantly affect corn borers (PubMed:25531527). In
CC contrast, NAL reduces the weight gain of corn borer larvae without
CC changing larval feeding behavior, indicating that its effect is due to
CC metabolic toxicity. N-formylloline, NAL, and NML are almost as potent
CC as nicotine in insecticidal activity against green bugs
CC (PubMed:25531527). {ECO:0000305|PubMed:25531527}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; AY723749; AAV68708.1; -; Genomic_DNA.
DR AlphaFoldDB; Q5MNI1; -.
DR SMR; Q5MNI1; -.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
PE 2: Evidence at transcript level;
KW Alkaloid metabolism; Glycoprotein; Heme; Iron; Membrane; Metal-binding;
KW Monooxygenase; Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..496
FT /note="Cytochrome P450 monooxygenase lolP1"
FT /id="PRO_0000444363"
FT TRANSMEM 10..30
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 443
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250|UniProtKB:P04798"
FT CARBOHYD 252
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
SQ SEQUENCE 496 AA; 54981 MW; 7207902A1AED974F CRC64;
MDLTQFNTAG IVWPTVAAIA ISYILLSSFL SWYRLRHIPG PFLASISSLW NVLNIVTGRT
SPVLEKLPGK YGPLVRTGPN YVLTDDAEIL RHVNGARSTY PRNGWYEGFK VDEHDHMGSH
IDTSVHDAIK SKVIGGYNGK DGIDLEGAIG SQVKTLVSEI RRRHLGQPVD FSRLMRQMAL
DAITAVAFGE ALGFLTAEDG DVFGYVSAVD KMLTYLTLAS DLPVVRSVVR SRRMAPAVRC
VLAYTGIGRM LNHTRRVVAE RYAADDPGKG DMTASFIRKG LTQIECEGES HLQLIAGADT
AVTVLRSTLL YIMTTPRVYT RLKAEIKAAV DAGEVVEVIT MAQAQRLPYL QAVVLEGFRM
RPAVVYGHFK SVPAGGDTLP NGVRLPAGTA IAPNYIALTR RADVYGADVD LFRPERFLDA
EPAKRHEMER AMDLNFGLGR WQCAGRNIAL MEMNKVFFEL LRHFDLQILY PGKAWDEYTG
VVYSQHNMWV QITESS
