KSA_ARATH
ID KSA_ARATH Reviewed; 802 AA.
AC Q38802;
DT 26-APR-2005, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 156.
DE RecName: Full=Ent-copalyl diphosphate synthase, chloroplastic;
DE Short=AtCPS {ECO:0000303|PubMed:17457817};
DE Short=Ent-CDP synthase;
DE EC=5.5.1.13 {ECO:0000269|PubMed:17457817, ECO:0000269|PubMed:20430888};
DE AltName: Full=Ent-kaurene synthase A;
DE Short=KSA;
DE AltName: Full=Protein GA REQUIRING 1;
DE Flags: Precursor;
GN Name=GA1; Synonyms=ABC33, CPS, CPS1, TPSGA1; OrderedLocusNames=At4g02780;
GN ORFNames=T5J8.9;
OS Arabidopsis thaliana (Mouse-ear cress).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; malvids; Brassicales; Brassicaceae; Camelineae; Arabidopsis.
OX NCBI_TaxID=3702;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA].
RC STRAIN=cv. Columbia;
RX PubMed=7994182; DOI=10.2307/3869986;
RA Sun T.-P., Kamiya Y.;
RT "The Arabidopsis GA1 locus encodes the cyclase ent-kaurene synthetase A of
RT gibberellin biosynthesis.";
RL Plant Cell 6:1509-1518(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=cv. Columbia;
RX PubMed=10617198; DOI=10.1038/47134;
RA Mayer K.F.X., Schueller C., Wambutt R., Murphy G., Volckaert G., Pohl T.,
RA Duesterhoeft A., Stiekema W., Entian K.-D., Terryn N., Harris B.,
RA Ansorge W., Brandt P., Grivell L.A., Rieger M., Weichselgartner M.,
RA de Simone V., Obermaier B., Mache R., Mueller M., Kreis M., Delseny M.,
RA Puigdomenech P., Watson M., Schmidtheini T., Reichert B., Portetelle D.,
RA Perez-Alonso M., Boutry M., Bancroft I., Vos P., Hoheisel J.,
RA Zimmermann W., Wedler H., Ridley P., Langham S.-A., McCullagh B.,
RA Bilham L., Robben J., van der Schueren J., Grymonprez B., Chuang Y.-J.,
RA Vandenbussche F., Braeken M., Weltjens I., Voet M., Bastiaens I., Aert R.,
RA Defoor E., Weitzenegger T., Bothe G., Ramsperger U., Hilbert H., Braun M.,
RA Holzer E., Brandt A., Peters S., van Staveren M., Dirkse W., Mooijman P.,
RA Klein Lankhorst R., Rose M., Hauf J., Koetter P., Berneiser S., Hempel S.,
RA Feldpausch M., Lamberth S., Van den Daele H., De Keyser A., Buysshaert C.,
RA Gielen J., Villarroel R., De Clercq R., van Montagu M., Rogers J.,
RA Cronin A., Quail M.A., Bray-Allen S., Clark L., Doggett J., Hall S.,
RA Kay M., Lennard N., McLay K., Mayes R., Pettett A., Rajandream M.A.,
RA Lyne M., Benes V., Rechmann S., Borkova D., Bloecker H., Scharfe M.,
RA Grimm M., Loehnert T.-H., Dose S., de Haan M., Maarse A.C., Schaefer M.,
RA Mueller-Auer S., Gabel C., Fuchs M., Fartmann B., Granderath K., Dauner D.,
RA Herzl A., Neumann S., Argiriou A., Vitale D., Liguori R., Piravandi E.,
RA Massenet O., Quigley F., Clabauld G., Muendlein A., Felber R., Schnabl S.,
RA Hiller R., Schmidt W., Lecharny A., Aubourg S., Chefdor F., Cooke R.,
RA Berger C., Monfort A., Casacuberta E., Gibbons T., Weber N., Vandenbol M.,
RA Bargues M., Terol J., Torres A., Perez-Perez A., Purnelle B., Bent E.,
RA Johnson S., Tacon D., Jesse T., Heijnen L., Schwarz S., Scholler P.,
RA Heber S., Francs P., Bielke C., Frishman D., Haase D., Lemcke K.,
RA Mewes H.-W., Stocker S., Zaccaria P., Bevan M., Wilson R.K.,
RA de la Bastide M., Habermann K., Parnell L., Dedhia N., Gnoj L., Schutz K.,
RA Huang E., Spiegel L., Sekhon M., Murray J., Sheet P., Cordes M.,
RA Abu-Threideh J., Stoneking T., Kalicki J., Graves T., Harmon G.,
RA Edwards J., Latreille P., Courtney L., Cloud J., Abbott A., Scott K.,
RA Johnson D., Minx P., Bentley D., Fulton B., Miller N., Greco T., Kemp K.,
RA Kramer J., Fulton L., Mardis E., Dante M., Pepin K., Hillier L.W.,
RA Nelson J., Spieth J., Ryan E., Andrews S., Geisel C., Layman D., Du H.,
RA Ali J., Berghoff A., Jones K., Drone K., Cotton M., Joshu C., Antonoiu B.,
RA Zidanic M., Strong C., Sun H., Lamar B., Yordan C., Ma P., Zhong J.,
RA Preston R., Vil D., Shekher M., Matero A., Shah R., Swaby I.K.,
RA O'Shaughnessy A., Rodriguez M., Hoffman J., Till S., Granat S., Shohdy N.,
RA Hasegawa A., Hameed A., Lodhi M., Johnson A., Chen E., Marra M.A.,
RA Martienssen R., McCombie W.R.;
RT "Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana.";
RL Nature 402:769-777(1999).
RN [3]
RP GENOME REANNOTATION.
RC STRAIN=cv. Columbia;
RX PubMed=27862469; DOI=10.1111/tpj.13415;
RA Cheng C.Y., Krishnakumar V., Chan A.P., Thibaud-Nissen F., Schobel S.,
RA Town C.D.;
RT "Araport11: a complete reannotation of the Arabidopsis thaliana reference
RT genome.";
RL Plant J. 89:789-804(2017).
RN [4]
RP GENE FAMILY, AND NOMENCLATURE.
RX PubMed=12207221; DOI=10.1007/s00438-002-0709-y;
RA Aubourg S., Lecharny A., Bohlmann J.;
RT "Genomic analysis of the terpenoid synthase (AtTPS) gene family of
RT Arabidopsis thaliana.";
RL Mol. Genet. Genomics 267:730-745(2002).
RN [5]
RP TISSUE SPECIFICITY.
RX PubMed=12566586; DOI=10.1105/tpc.007989;
RA Chen F., Tholl D., D'Auria J.C., Farooq A., Pichersky E., Gershenzon J.;
RT "Biosynthesis and emission of terpenoid volatiles from Arabidopsis
RT flowers.";
RL Plant Cell 15:481-494(2003).
RN [6]
RP GENE FAMILY.
RX PubMed=12777052; DOI=10.1023/a:1023005504702;
RA Lange B.M., Ghassemian M.;
RT "Genome organization in Arabidopsis thaliana: a survey for genes involved
RT in isoprenoid and chlorophyll metabolism.";
RL Plant Mol. Biol. 51:925-948(2003).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY, AND
RP MUTAGENESIS OF ASP-379 AND ASP-380.
RX PubMed=17457817; DOI=10.1002/cbic.200700045;
RA Prisic S., Xu J., Coates R.M., Peters R.J.;
RT "Probing the role of the DXDD motif in Class II diterpene cyclases.";
RL ChemBioChem 8:869-874(2007).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP ACTIVITY REGULATION, PATHWAY, AND MUTAGENESIS OF HIS-331 AND ASP-377.
RX PubMed=20430888; DOI=10.1074/jbc.m110.123307;
RA Mann F.M., Prisic S., Davenport E.K., Determan M.K., Coates R.M.,
RA Peters R.J.;
RT "A single residue switch for Mg(2+)-dependent inhibition characterizes
RT plant class II diterpene cyclases from primary and secondary metabolism.";
RL J. Biol. Chem. 285:20558-20563(2010).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (2.25 ANGSTROMS) OF 85-802 IN COMPLEX WITH SUBSTRATE
RP ANALOGS.
RX PubMed=21602811; DOI=10.1038/nchembio.578;
RA Koeksal M., Hu H., Coates R.M., Peters R.J., Christianson D.W.;
RT "Structure and mechanism of the diterpene cyclase ent-copalyl diphosphate
RT synthase.";
RL Nat. Chem. Biol. 7:431-433(2011).
CC -!- FUNCTION: Catalyzes the conversion of geranylgeranyl diphosphate to the
CC gibberellin precursor ent-copalyl diphosphate.
CC {ECO:0000269|PubMed:17457817, ECO:0000269|PubMed:20430888,
CC ECO:0000305|PubMed:7994182}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E,10E)-geranylgeranyl diphosphate = ent-copalyl
CC diphosphate; Xref=Rhea:RHEA:14841, ChEBI:CHEBI:58553,
CC ChEBI:CHEBI:58756; EC=5.5.1.13;
CC Evidence={ECO:0000269|PubMed:17457817, ECO:0000269|PubMed:20430888};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:14842;
CC Evidence={ECO:0000269|PubMed:17457817, ECO:0000269|PubMed:20430888};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:20430888};
CC -!- ACTIVITY REGULATION: Inhibited by high concentrations of magnesium.
CC {ECO:0000269|PubMed:20430888}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.9 uM for GGPP (at pH 7.75 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:17457817};
CC KM=3 uM for GGPP {ECO:0000269|PubMed:20430888};
CC -!- PATHWAY: Plant hormone biosynthesis; gibberellin biosynthesis.
CC {ECO:0000269|PubMed:17457817, ECO:0000269|PubMed:20430888}.
CC -!- SUBCELLULAR LOCATION: Plastid, chloroplast.
CC -!- TISSUE SPECIFICITY: Expressed in roots, leaves, flowers and also in
CC siliques. {ECO:0000269|PubMed:12566586}.
CC -!- DOMAIN: The Asp-Xaa-Asp-Asp (DXDD) motif is important for the catalytic
CC activity through binding to Mg(2+).
CC -!- PTM: The N-terminus is blocked.
CC -!- SIMILARITY: Belongs to the terpene synthase family. Tpsc subfamily.
CC {ECO:0000305}.
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DR EMBL; U11034; AAA53632.1; -; mRNA.
DR EMBL; AC004044; AAD15334.1; -; Genomic_DNA.
DR EMBL; AL161495; CAB77763.1; -; Genomic_DNA.
DR EMBL; CP002687; AEE82229.1; -; Genomic_DNA.
DR PIR; D85035; D85035.
DR RefSeq; NP_192187.1; NM_116512.2.
DR PDB; 3PYA; X-ray; 2.25 A; A=85-802.
DR PDB; 3PYB; X-ray; 2.76 A; A/B/C=85-802.
DR PDB; 4LIX; X-ray; 1.55 A; A=85-802.
DR PDBsum; 3PYA; -.
DR PDBsum; 3PYB; -.
DR PDBsum; 4LIX; -.
DR AlphaFoldDB; Q38802; -.
DR SMR; Q38802; -.
DR STRING; 3702.AT4G02780.1; -.
DR PaxDb; Q38802; -.
DR PRIDE; Q38802; -.
DR EnsemblPlants; AT4G02780.1; AT4G02780.1; AT4G02780.
DR GeneID; 828182; -.
DR Gramene; AT4G02780.1; AT4G02780.1; AT4G02780.
DR KEGG; ath:AT4G02780; -.
DR Araport; AT4G02780; -.
DR TAIR; locus:2140260; AT4G02780.
DR eggNOG; ENOG502QQN6; Eukaryota.
DR HOGENOM; CLU_003125_3_2_1; -.
DR InParanoid; Q38802; -.
DR OMA; RDKWVIA; -.
DR OrthoDB; 700680at2759; -.
DR PhylomeDB; Q38802; -.
DR BioCyc; ARA:AT4G02780-MON; -.
DR BioCyc; MetaCyc:AT4G02780-MON; -.
DR BRENDA; 5.5.1.13; 399.
DR UniPathway; UPA00390; -.
DR EvolutionaryTrace; Q38802; -.
DR PRO; PR:Q38802; -.
DR Proteomes; UP000006548; Chromosome 4.
DR ExpressionAtlas; Q38802; baseline and differential.
DR Genevisible; Q38802; AT.
DR GO; GO:0009507; C:chloroplast; IDA:TAIR.
DR GO; GO:0009905; F:ent-copalyl diphosphate synthase activity; IDA:TAIR.
DR GO; GO:0000287; F:magnesium ion binding; IDA:TAIR.
DR GO; GO:0010333; F:terpene synthase activity; IBA:GO_Central.
DR GO; GO:0016102; P:diterpenoid biosynthetic process; IBA:GO_Central.
DR GO; GO:0009740; P:gibberellic acid mediated signaling pathway; TAS:TAIR.
DR GO; GO:0009686; P:gibberellin biosynthetic process; IDA:TAIR.
DR CDD; cd00684; Terpene_cyclase_plant_C1; 1.
DR Gene3D; 1.10.600.10; -; 1.
DR Gene3D; 1.50.10.130; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR044814; Terpene_cyclase_plant_C1.
DR InterPro; IPR001906; Terpene_synth_N.
DR InterPro; IPR036965; Terpene_synth_N_sf.
DR InterPro; IPR005630; Terpene_synthase_metal-bd.
DR InterPro; IPR008930; Terpenoid_cyclase/PrenylTrfase.
DR Pfam; PF01397; Terpene_synth; 1.
DR Pfam; PF03936; Terpene_synth_C; 1.
DR SUPFAM; SSF48239; SSF48239; 2.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Chloroplast; Isomerase; Magnesium; Metal-binding; Plastid;
KW Reference proteome; Transit peptide.
FT TRANSIT 1..60
FT /note="Chloroplast"
FT /evidence="ECO:0000255"
FT CHAIN 61..802
FT /note="Ent-copalyl diphosphate synthase, chloroplastic"
FT /id="PRO_0000033623"
FT MOTIF 377..380
FT /note="DXDD motif"
FT BINDING 245
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21602811,
FT ECO:0007744|PDB:3PYA"
FT BINDING 377
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:C7BKP9"
FT BINDING 379
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:C7BKP9"
FT BINDING 463
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21602811,
FT ECO:0007744|PDB:3PYA"
FT MUTAGEN 331
FT /note="H->A: Reduced activity, but no effect on substrate
FT binding. Increased inhibition by Mg(2+)."
FT /evidence="ECO:0000269|PubMed:20430888"
FT MUTAGEN 331
FT /note="H->R: Reduced catalytic activity, but loss of
FT inhibition by Mg(2+)."
FT /evidence="ECO:0000269|PubMed:20430888"
FT MUTAGEN 377
FT /note="D->A: Reduced catalytic activity, but no effect on
FT substrate binding. Reduced inhibition by Mg(2+)."
FT /evidence="ECO:0000269|PubMed:20430888"
FT MUTAGEN 379
FT /note="D->A: Loss of function."
FT /evidence="ECO:0000269|PubMed:17457817"
FT MUTAGEN 380
FT /note="D->A: Strongly reduced activity. Reduced inhibition
FT by Mg(2+)."
FT /evidence="ECO:0000269|PubMed:17457817"
FT HELIX 92..106
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 117..124
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 129..134
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 136..145
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 155..157
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 160..176
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 181..194
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 195..197
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 203..205
FT /evidence="ECO:0007829|PDB:3PYB"
FT HELIX 210..223
FT /evidence="ECO:0007829|PDB:4LIX"
FT TURN 232..234
FT /evidence="ECO:0007829|PDB:3PYB"
FT HELIX 235..248
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 251..254
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 260..267
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 273..276
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 277..279
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 284..286
FT /evidence="ECO:0007829|PDB:3PYA"
FT HELIX 290..300
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 303..315
FT /evidence="ECO:0007829|PDB:4LIX"
FT TURN 316..318
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 322..324
FT /evidence="ECO:0007829|PDB:3PYB"
FT HELIX 327..340
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 344..347
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 348..361
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 369..372
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 378..390
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 397..403
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 420..430
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 438..456
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 463..466
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 470..479
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 482..484
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 487..497
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 500..502
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 504..511
FT /evidence="ECO:0007829|PDB:4LIX"
FT TURN 514..516
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 519..549
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 552..555
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 559..570
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 576..578
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 579..599
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 603..615
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 640..664
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 669..685
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 686..688
FT /evidence="ECO:0007829|PDB:3PYB"
FT HELIX 691..701
FT /evidence="ECO:0007829|PDB:4LIX"
FT STRAND 703..705
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 708..712
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 714..726
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 742..760
FT /evidence="ECO:0007829|PDB:4LIX"
FT TURN 764..766
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 767..786
FT /evidence="ECO:0007829|PDB:4LIX"
FT HELIX 791..799
FT /evidence="ECO:0007829|PDB:4LIX"
SQ SEQUENCE 802 AA; 93014 MW; B2815AB0C06414F6 CRC64;
MSLQYHVLNS IPSTTFLSST KTTISSSFLT ISGSPLNVAR DKSRSGSIHC SKLRTQEYIN
SQEVQHDLPL IHEWQQLQGE DAPQISVGSN SNAFKEAVKS VKTILRNLTD GEITISAYDT
AWVALIDAGD KTPAFPSAVK WIAENQLSDG SWGDAYLFSY HDRLINTLAC VVALRSWNLF
PHQCNKGITF FRENIGKLED ENDEHMPIGF EVAFPSLLEI ARGINIDVPY DSPVLKDIYA
KKELKLTRIP KEIMHKIPTT LLHSLEGMRD LDWEKLLKLQ SQDGSFLFSP SSTAFAFMQT
RDSNCLEYLR NAVKRFNGGV PNVFPVDLFE HIWIVDRLQR LGISRYFEEE IKECLDYVHR
YWTDNGICWA RCSHVQDIDD TAMAFRLLRQ HGYQVSADVF KNFEKEGEFF CFVGQSNQAV
TGMFNLYRAS QLAFPREEIL KNAKEFSYNY LLEKREREEL IDKWIIMKDL PGEIGFALEI
PWYASLPRVE TRFYIDQYGG ENDVWIGKTL YRMPYVNNNG YLELAKQDYN NCQAQHQLEW
DIFQKWYEEN RLSEWGVRRS ELLECYYLAA ATIFESERSH ERMVWAKSSV LVKAISSSFG
ESSDSRRSFS DQFHEYIANA RRSDHHFNDR NMRLDRPGSV QASRLAGVLI GTLNQMSFDL
FMSHGRDVNN LLYLSWGDWM EKWKLYGDEG EGELMVKMII LMKNNDLTNF FTHTHFVRLA
EIINRICLPR QYLKARRNDE KEKTIKSMEK EMGKMVELAL SESDTFRDVS ITFLDVAKAF
YYFALCGDHL QTHISKVLFQ KV
