PAM_TAXWC
ID PAM_TAXWC Reviewed; 687 AA.
AC Q68G84;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 11-OCT-2004, sequence version 1.
DT 03-AUG-2022, entry version 104.
DE RecName: Full=Phenylalanine aminomutase (L-beta-phenylalanine forming);
DE EC=5.4.3.10 {ECO:0000269|PubMed:15878763, ECO:0000269|PubMed:22113970};
DE AltName: Full=Phenylalanine ammonia-lyase;
DE EC=4.3.1.24 {ECO:0000250|UniProtKB:Q6GZ04};
GN Name=pam;
OS Taxus wallichiana var. chinensis (Chinese yew) (Taxus chinensis).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Pinopsida; Pinidae; Conifers II; Cupressales; Taxaceae;
OC Taxus.
OX NCBI_TaxID=29808;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PARTIAL PROTEIN SEQUENCE, CATALYTIC ACTIVITY,
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=15878763; DOI=10.1016/j.abb.2005.04.012;
RA Steele C.L., Chen Y., Dougherty B.A., Li W., Hofstead S., Lam K.S.,
RA Xing Z., Chiang S.J.;
RT "Purification, cloning, and functional expression of phenylalanine
RT aminomutase: the first committed step in Taxol side-chain biosynthesis.";
RL Arch. Biochem. Biophys. 438:1-10(2005).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.18 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, MUTAGENESIS OF ASN-231;
RP GLN-319; TYR-322; ARG-325; ASN-355 AND PHE-371, ACTIVE SITE, AND PTM.
RX PubMed=22113970; DOI=10.1002/anie.201106372;
RA Wu B., Szymanski W., Wybenga G.G., Heberling M.M., Bartsch S.,
RA de Wildeman S., Poelarends G.J., Feringa B.L., Dijkstra B.W., Janssen D.B.;
RT "Mechanism-inspired engineering of phenylalanine aminomutase for enhanced
RT beta-regioselective asymmetric amination of cinnamates.";
RL Angew. Chem. Int. Ed. 51:482-486(2012).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEX WITH TRANS-CINNAMATE,
RP FUNCTION, ACTIVE SITE, PTM, DEHYDRATION AT SER-176, SUBUNIT, AND
RP MUTAGENESIS OF TYR-80; ASN-231 AND TYR-322.
RX PubMed=24786474; DOI=10.1021/bi500187a;
RA Wybenga G.G., Szymanski W., Wu B., Feringa B.L., Janssen D.B.,
RA Dijkstra B.W.;
RT "Structural investigations into the stereochemistry and activity of a
RT phenylalanine-2,3-aminomutase from Taxus chinensis.";
RL Biochemistry 53:3187-3198(2014).
CC -!- FUNCTION: Phenylalanine aminomutase that catalyzes the rearrangement of
CC L-phenylalanine to R-beta-phenylalanine. Catalyzes the first committed
CC step in the biosynthesis of the side chain of the alkaloid taxol
CC (paclitaxel), a widely-used compound with antitumor activity. Has also
CC low phenylalanine ammonia-lyase activity and can catalyze the amination
CC of trans-cinnamate. {ECO:0000269|PubMed:15878763,
CC ECO:0000269|PubMed:22113970, ECO:0000269|PubMed:24786474}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-phenylalanine = L-beta-phenylalanine; Xref=Rhea:RHEA:34395,
CC ChEBI:CHEBI:58095, ChEBI:CHEBI:67158; EC=5.4.3.10;
CC Evidence={ECO:0000269|PubMed:15878763, ECO:0000269|PubMed:22113970};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-phenylalanine = (E)-cinnamate + NH4(+);
CC Xref=Rhea:RHEA:21384, ChEBI:CHEBI:15669, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:58095; EC=4.3.1.24;
CC Evidence={ECO:0000250|UniProtKB:Q6GZ04};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.1 mM for L-phenylalanine {ECO:0000269|PubMed:15878763,
CC ECO:0000269|PubMed:22113970};
CC Vmax=110 umol/min/mg enzyme with L-phenylalanine as substrate
CC {ECO:0000269|PubMed:15878763, ECO:0000269|PubMed:22113970};
CC pH dependence:
CC Optimum pH is 7.5-8.0. {ECO:0000269|PubMed:15878763,
CC ECO:0000269|PubMed:22113970};
CC -!- PATHWAY: Alkaloid biosynthesis; taxol biosynthesis.
CC -!- PATHWAY: Phenylpropanoid metabolism; trans-cinnamate biosynthesis;
CC trans-cinnamate from L-phenylalanine: step 1/1. {ECO:0000305}.
CC -!- SUBUNIT: Homodimer (PubMed:15878763). Homotetramer, dimer of dimers
CC (PubMed:24786474). {ECO:0000269|PubMed:15878763,
CC ECO:0000269|PubMed:24786474}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC -!- PTM: Contains an active site 4-methylidene-imidazol-5-one (MIO), which
CC is formed autocatalytically by cyclization and dehydration of residues
CC Ala-Ser-Gly. {ECO:0000269|PubMed:24786474}.
CC -!- BIOTECHNOLOGY: Could be used for the stereoselective biosynthesis of
CC beta-amino acids via amination of cinnamic acid derivatives.
CC {ECO:0000269|PubMed:22113970}.
CC -!- SIMILARITY: Belongs to the PAL/histidase family. {ECO:0000305}.
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DR EMBL; AY724735; AAU01182.1; -; mRNA.
DR PDB; 2YII; X-ray; 2.18 A; A/B/C/D=1-687.
DR PDB; 4BAA; X-ray; 2.50 A; A/B/C/D=1-687.
DR PDB; 4BAB; X-ray; 2.56 A; A/B/C/D=1-687.
DR PDB; 4C5R; X-ray; 2.14 A; A/B/C/D=1-687.
DR PDB; 4C5S; X-ray; 1.85 A; A/B/C/D=1-687.
DR PDB; 4C5U; X-ray; 2.19 A; A/B/C/D=1-687.
DR PDB; 4C6G; X-ray; 2.10 A; A/B/C/D=1-687.
DR PDB; 4CQ5; X-ray; 1.90 A; A/B/C/D=1-687.
DR PDB; 4V2Q; X-ray; 1.95 A; A/B=1-687.
DR PDB; 4V2R; X-ray; 2.20 A; A/B=1-687.
DR PDBsum; 2YII; -.
DR PDBsum; 4BAA; -.
DR PDBsum; 4BAB; -.
DR PDBsum; 4C5R; -.
DR PDBsum; 4C5S; -.
DR PDBsum; 4C5U; -.
DR PDBsum; 4C6G; -.
DR PDBsum; 4CQ5; -.
DR PDBsum; 4V2Q; -.
DR PDBsum; 4V2R; -.
DR AlphaFoldDB; Q68G84; -.
DR SMR; Q68G84; -.
DR BRENDA; 5.4.3.10; 9720.
DR BRENDA; 5.4.3.11; 9720.
DR UniPathway; UPA00713; UER00725.
DR UniPathway; UPA00842; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0016869; F:intramolecular transferase activity, transferring amino groups; IDA:UniProtKB.
DR GO; GO:0045548; F:phenylalanine ammonia-lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0009821; P:alkaloid biosynthetic process; TAS:UniProtKB.
DR GO; GO:0009800; P:cinnamic acid biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0006559; P:L-phenylalanine catabolic process; IEA:UniProtKB-KW.
DR GO; GO:0006558; P:L-phenylalanine metabolic process; IDA:UniProtKB.
DR GO; GO:0042617; P:paclitaxel biosynthetic process; TAS:UniProtKB.
DR GO; GO:0051289; P:protein homotetramerization; IDA:UniProtKB.
DR CDD; cd00332; PAL-HAL; 1.
DR Gene3D; 1.10.274.20; -; 1.
DR Gene3D; 1.10.275.10; -; 1.
DR InterPro; IPR001106; Aromatic_Lyase.
DR InterPro; IPR024083; Fumarase/histidase_N.
DR InterPro; IPR008948; L-Aspartase-like.
DR InterPro; IPR022313; Phe/His_NH3-lyase_AS.
DR InterPro; IPR005922; Phe_NH3-lyase.
DR InterPro; IPR023144; Phe_NH3-lyase_shielding_dom_sf.
DR InterPro; IPR031008; Taxol_Phe_23mut.
DR PANTHER; PTHR10362; PTHR10362; 1.
DR Pfam; PF00221; Lyase_aromatic; 1.
DR SUPFAM; SSF48557; SSF48557; 1.
DR TIGRFAMs; TIGR01226; phe_am_lyase; 1.
DR TIGRFAMs; TIGR04473; taxol_Phe_23mut; 1.
DR PROSITE; PS00488; PAL_HISTIDASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Cytoplasm; Direct protein sequencing;
KW Isomerase; Lyase; Phenylalanine catabolism; Phenylpropanoid metabolism;
KW Taxol biosynthesis.
FT CHAIN 1..687
FT /note="Phenylalanine aminomutase (L-beta-phenylalanine
FT forming)"
FT /id="PRO_0000429970"
FT ACT_SITE 80
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000269|PubMed:22113970,
FT ECO:0000269|PubMed:24786474"
FT BINDING 231
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24786474,
FT ECO:0007744|PDB:4CQ5"
FT BINDING 319
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24786474,
FT ECO:0007744|PDB:4CQ5"
FT BINDING 325
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24786474,
FT ECO:0007744|PDB:4CQ5"
FT BINDING 355
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24786474,
FT ECO:0007744|PDB:4CQ5"
FT BINDING 458
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:24786474,
FT ECO:0007744|PDB:4CQ5"
FT MOD_RES 176
FT /note="2,3-didehydroalanine (Ser)"
FT /evidence="ECO:0000269|PubMed:24786474"
FT CROSSLNK 175..177
FT /note="5-imidazolinone (Ala-Gly)"
FT /evidence="ECO:0000269|PubMed:24786474"
FT MUTAGEN 80
FT /note="Y->A,F: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:24786474"
FT MUTAGEN 231
FT /note="N->A: Abolishes the formation of the MIO cofactor
FT and thereby abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:22113970,
FT ECO:0000269|PubMed:24786474"
FT MUTAGEN 231
FT /note="N->X: Abolishes enzyme activity; when associated
FT with X-355."
FT /evidence="ECO:0000269|PubMed:22113970,
FT ECO:0000269|PubMed:24786474"
FT MUTAGEN 319
FT /note="Q->M: Increases deamination activity with beta-Phe.
FT Increases beta-regioselectivity in the amination of
FT cinnamate. Abolishes enzyme activity; when associated with
FT K-325."
FT /evidence="ECO:0000269|PubMed:22113970"
FT MUTAGEN 322
FT /note="Y->A: Abolishes the formation of the MIO cofactor
FT and thereby abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:22113970,
FT ECO:0000269|PubMed:24786474"
FT MUTAGEN 322
FT /note="Y->X: Abolishes enzyme activity; when associated
FT with X-371."
FT /evidence="ECO:0000269|PubMed:22113970,
FT ECO:0000269|PubMed:24786474"
FT MUTAGEN 325
FT /note="R->K: Increases deamination activity with beta-Phe.
FT Increases beta-regioselectivity in the amination of
FT cinnamate. Abolishes enzyme activity; when associated with
FT M-319."
FT /evidence="ECO:0000269|PubMed:22113970"
FT MUTAGEN 355
FT /note="N->X: Abolishes enzyme activity; when associated
FT with X-231."
FT /evidence="ECO:0000269|PubMed:22113970"
FT MUTAGEN 371
FT /note="F->X: Abolishes enzyme activity; when associated
FT with X-322."
FT /evidence="ECO:0000269|PubMed:22113970"
FT HELIX 10..23
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 26..33
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 37..45
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 50..53
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 55..74
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 80..82
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 87..89
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 97..108
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 126..140
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 149..160
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 163..165
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 174..176
FT /evidence="ECO:0007829|PDB:4C6G"
FT HELIX 179..189
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 196..199
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 200..202
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 203..206
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 207..213
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 225..230
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 231..233
FT /evidence="ECO:0007829|PDB:4C6G"
FT HELIX 234..265
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 269..272
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 274..278
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 283..296
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 300..310
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 313..315
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 322..325
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 327..349
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 355..359
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 360..362
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 364..366
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 374..402
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 404..406
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 407..409
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 412..414
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 420..422
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 427..443
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 448..450
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 455..458
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 459..461
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 465..517
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 522..534
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 537..539
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 540..542
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 550..565
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 575..603
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 613..615
FT /evidence="ECO:0007829|PDB:4V2R"
FT HELIX 619..622
FT /evidence="ECO:0007829|PDB:4C5S"
FT HELIX 626..633
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 634..637
FT /evidence="ECO:0007829|PDB:4C5S"
FT STRAND 642..644
FT /evidence="ECO:0007829|PDB:4BAA"
FT HELIX 649..661
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 662..665
FT /evidence="ECO:0007829|PDB:4CQ5"
FT HELIX 666..672
FT /evidence="ECO:0007829|PDB:4C5S"
FT TURN 673..675
FT /evidence="ECO:0007829|PDB:4C5S"
SQ SEQUENCE 687 AA; 75332 MW; D9BC13C7C689A421 CRC64;
MGFAVESRSH VKDILGLINA FNEVKKITVD GTTPITVAHV AALARRHDVK VALEAEQCRA
RVETCSSWVQ RKAEDGADIY GVTTGFGACS SRRTNRLSEL QESLIRCLLA GVFTKGCAPS
VDELPATATR SAMLLRLNSF TYGCSGIRWE VMEALEKLLN SNVSPKVPLR GSVSASGDLI
PLAYIAGLLI GKPSVIARIG DDVEVPAPEA LSRVGLRPFK LQAKEGLALV NGTSFATAVA
STVMYDANVL LLLVETLCGM FCEVIFGREE FAHPLIHKVK PHPGQIESAE LLEWLLRSSP
FQELSREYYS IDKLKKPKQD RYALRSSPQW LAPLVQTIRD ATTTVETEVN SANDNPIIDH
ANDRALHGAN FQGSAVGFYM DYVRIAVAGL GKLLFAQFTE LMIEYYSNGL PGNLSLGPDL
SVDYGLKGLD IAMAAYSSEL QYLANPVTTH VHSAEQHNQD INSLALISAR KTEEALDILK
LMIASHLTAM CQAVDLRQLE EALVKVVENV VSTLADECGL PNDTKARLLY VAKAVPVYTY
LESPCDPTLP LLLGLKQSCF DTILALHKKD GIETDTLVDR LAEFEKRLSD RLENEMTAVR
VLYEKKGHKT ADNNDALVRI QGSKFLPFYR FVREELDTGV MSARREQTPQ EDVQKVFDAI
ADGRITVPLL HCLQGFLGQP NGCANGV
