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Protein

CAD protein

Gene

CAD

Organism
Homo sapiens (Human)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

This protein is a "fusion" protein encoding four enzymatic activities of the pyrimidine pathway (GATase, CPSase, ATCase and DHOase).1 Publication

Catalytic activityi

2 ATP + L-glutamine + HCO3- + H2O = 2 ADP + phosphate + L-glutamate + carbamoyl phosphate.1 Publication
Carbamoyl phosphate + L-aspartate = phosphate + N-carbamoyl-L-aspartate.1 Publication
(S)-dihydroorotate + H2O = N-carbamoyl-L-aspartate.1 Publication

Cofactori

Zn2+2 PublicationsNote: Binds 3 Zn2+ ions per subunit (for dihydroorotase activity).2 Publications

Enzyme regulationi

Allosterically regulated and controlled by phosphorylation. 5-phosphoribose 1-diphosphate (PRPP) is an activator while UMP and UTP are inhibitors of the CPSase reaction.1 Publication

Kineticsi

  1. KM=28 µM for dihydroorotate1 Publication
  2. KM=241 µM for N-carbamoyl-L-aspartate1 Publication

    Pathway:iUMP biosynthesis via de novo pathway

    This protein is involved in step 1, 2 and 3 of the subpathway that synthesizes (S)-dihydroorotate from bicarbonate.
    Proteins known to be involved in the 3 steps of the subpathway in this organism are:
    1. CAD protein (CAD)
    2. CAD protein (CAD)
    3. CAD protein (CAD)
    This subpathway is part of the pathway UMP biosynthesis via de novo pathway, which is itself part of Pyrimidine metabolism.
    View all proteins of this organism that are known to be involved in the subpathway that synthesizes (S)-dihydroorotate from bicarbonate, the pathway UMP biosynthesis via de novo pathway and in Pyrimidine metabolism.

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Active sitei252 – 2521For GATase activityBy similarity
    Active sitei336 – 3361For GATase activityBy similarity
    Active sitei338 – 3381For GATase activityBy similarity
    Metal bindingi1471 – 14711Zinc 1; via tele nitrogen1 Publication
    Metal bindingi1471 – 14711Zinc 2; via pros nitrogen1 Publication
    Metal bindingi1473 – 14731Zinc 1; via tele nitrogen1 Publication
    Binding sitei1475 – 14751N-carbamoyl-L-aspartate
    Binding sitei1505 – 15051N-carbamoyl-L-aspartate
    Metal bindingi1556 – 15561Zinc 1; via carbamate group1 Publication
    Metal bindingi1556 – 15561Zinc 3; via carbamate group1 Publication
    Metal bindingi1590 – 15901Zinc 3; via pros nitrogen1 Publication
    Metal bindingi1613 – 16131Zinc 21 Publication
    Metal bindingi1614 – 16141Zinc 3; via tele nitrogen1 Publication
    Metal bindingi1637 – 16371Zinc 21 Publication
    Binding sitei1661 – 16611N-carbamoyl-L-aspartate; via amide nitrogen and carbonyl oxygen
    Metal bindingi1686 – 16861Zinc 11 Publication
    Binding sitei1686 – 16861N-carbamoyl-L-aspartate
    Binding sitei1690 – 16901N-carbamoyl-L-aspartate

    GO - Molecular functioni

    GO - Biological processi

    Keywords - Molecular functioni

    Hydrolase, Ligase, Transferase

    Keywords - Biological processi

    Pyrimidine biosynthesis

    Keywords - Ligandi

    ATP-binding, Metal-binding, Nucleotide-binding, Zinc

    Enzyme and pathway databases

    BioCyciMetaCyc:ENSG00000084774-MONOMER.
    BRENDAi3.5.2.3. 2681.
    ReactomeiREACT_21376. Pyrimidine biosynthesis.
    UniPathwayiUPA00070; UER00115.
    UPA00070; UER00116.
    UPA00070; UER00117.

    Protein family/group databases

    MEROPSiM38.972.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    CAD protein
    Including the following 3 domains:
    Glutamine-dependent carbamoyl-phosphate synthase (EC:6.3.5.5)
    Aspartate carbamoyltransferase (EC:2.1.3.2)
    Dihydroorotase (EC:3.5.2.3)
    Gene namesi
    Name:CAD
    OrganismiHomo sapiens (Human)
    Taxonomic identifieri9606 [NCBI]
    Taxonomic lineageiEukaryotaMetazoaChordataCraniataVertebrataEuteleostomiMammaliaEutheriaEuarchontogliresPrimatesHaplorrhiniCatarrhiniHominidaeHomo
    ProteomesiUP000005640 Componenti: Chromosome 2

    Organism-specific databases

    HGNCiHGNC:1424. CAD.

    Subcellular locationi

    GO - Cellular componenti

    • cell projection Source: BHF-UCL
    • cytoplasm Source: GO_Central
    • cytosol Source: BHF-UCL
    • extracellular exosome Source: UniProtKB
    • membrane Source: UniProtKB
    • neuronal cell body Source: BHF-UCL
    • nuclear matrix Source: BHF-UCL
    • nucleoplasm Source: HPA
    • nucleus Source: BHF-UCL
    • plasma membrane Source: HPA
    • protein complex Source: Ensembl
    • terminal bouton Source: BHF-UCL

    Keywords - Cellular componenti

    Cytoplasm, Nucleus

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi1471 – 14711H → A: No zinc-binding and no catalytic activity. 2 Publications
    Mutagenesisi1471 – 14711H → N: Abolishes dihydroorotase activity. 2 Publications
    Mutagenesisi1473 – 14731H → A: No zinc-binding and no catalytic activity. 1 Publication
    Mutagenesisi1512 – 15121D → N: No change in catalytic activity. 1 Publication
    Mutagenesisi1562 – 15621T → A: Abolishes dihydroorotase activity. 1 Publication
    Mutagenesisi1563 – 15631F → A: Abolishes dihydroorotase activity. 1 Publication
    Mutagenesisi1590 – 15901H → A: Abolishes dihydroorotase activity. 2 Publications
    Mutagenesisi1590 – 15901H → N: No catalytic activity. 2 Publications
    Mutagenesisi1613 – 16131C → S: Reduces dihydroorotase activity. 1 Publication
    Mutagenesisi1614 – 16141H → A: Abolishes dihydroorotase activity. 1 Publication
    Mutagenesisi1637 – 16371E → T: Abolishes dihydroorotase activity. 1 Publication
    Mutagenesisi1642 – 16421H → N: 11.5% of wild-type catalytic activity. 1 Publication
    Mutagenesisi1686 – 16861D → N: Abolishes dihydroorotase activity. 1 Publication
    Mutagenesisi1690 – 16901H → N: 3% of wild-type catalytic activity. 1 Publication
    Mutagenesisi1873 – 18731S → A: Abolishes PMA-induced Thr-456 phosphorylation. 1 Publication

    Organism-specific databases

    PharmGKBiPA26023.

    Chemistry

    DrugBankiDB00128. L-Aspartic Acid.
    DB00130. L-Glutamine.

    Polymorphism and mutation databases

    BioMutaiCAD.
    DMDMi50403731.

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Initiator methioninei1 – 11Removed4 Publications
    Chaini2 – 22252224CAD proteinPRO_0000199506Add
    BLAST

    Amino acid modifications

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Modified residuei2 – 21N-acetylalanine4 Publications
    Modified residuei456 – 4561Phosphothreonine; by MAPK11 Publication
    Modified residuei747 – 7471N6-acetyllysine1 Publication
    Modified residuei1406 – 14061Phosphoserine; by PKA2 Publications
    Modified residuei1411 – 14111N6-acetyllysine1 Publication
    Modified residuei1556 – 15561N6-carboxylysine1 Publication
    Modified residuei1859 – 18591Phosphoserine; by RPS6KB1 and PKA9 Publications
    Modified residuei1873 – 18731Phosphoserine; by PKC; in vitro1 Publication
    Modified residuei1884 – 18841Phosphothreonine2 Publications
    Modified residuei1900 – 19001Phosphoserine2 Publications

    Post-translational modificationi

    Activated by MAP kinase (Erk1/2) phosphorylation just prior to the S phase of the cell cycle, when the demand for pyrimidine nucleotides is greatest, and down-regulated as the cells emerge from S phase by protein kinase A (PKA) phosphorylation. Phosphorylation at Ser-1859 by RPS6KB1 downstream of MTOR promotes oligomerization and stimulates dihydroorotase activity. Phosphorylation at Ser-1406 reduces sensitivy to feedback inhibition by UTP.3 Publications

    Keywords - PTMi

    Acetylation, Phosphoprotein

    Proteomic databases

    MaxQBiP27708.
    PaxDbiP27708.
    PeptideAtlasiP27708.
    PRIDEiP27708.

    PTM databases

    PhosphoSiteiP27708.

    Miscellaneous databases

    PMAP-CutDBP27708.

    Expressioni

    Inductioni

    Transcriptionally repressed following hypoxia by HIF1A.1 Publication

    Gene expression databases

    BgeeiP27708.
    CleanExiHS_CAD.
    ExpressionAtlasiP27708. baseline and differential.
    GenevisibleiP27708. HS.

    Organism-specific databases

    HPAiCAB007781.
    HPA050746.
    HPA057266.

    Interactioni

    Subunit structurei

    Homohexamer.1 Publication

    Protein-protein interaction databases

    BioGridi107243. 44 interactions.
    DIPiDIP-39484N.
    IntActiP27708. 25 interactions.
    MINTiMINT-5000537.
    STRINGi9606.ENSP00000264705.

    Structurei

    Secondary structure

    1
    2225
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi1462 – 14654Combined sources
    Beta strandi1467 – 14726Combined sources
    Turni1476 – 14783Combined sources
    Turni1480 – 14823Combined sources
    Helixi1485 – 149410Combined sources
    Beta strandi1497 – 15026Combined sources
    Beta strandi1506 – 15083Combined sources
    Helixi1513 – 152614Combined sources
    Beta strandi1528 – 15336Combined sources
    Turni1546 – 15483Combined sources
    Helixi1549 – 15513Combined sources
    Beta strandi1555 – 15584Combined sources
    Beta strandi1560 – 15634Combined sources
    Turni1564 – 15663Combined sources
    Helixi1571 – 158010Combined sources
    Beta strandi1587 – 15904Combined sources
    Helixi1594 – 160512Combined sources
    Beta strandi1610 – 16123Combined sources
    Helixi1618 – 162912Combined sources
    Beta strandi1634 – 16385Combined sources
    Helixi1640 – 16445Combined sources
    Helixi1647 – 16493Combined sources
    Helixi1650 – 16578Combined sources
    Helixi1667 – 16759Combined sources
    Helixi1677 – 16793Combined sources
    Helixi1692 – 16954Combined sources
    Beta strandi1697 – 16993Combined sources
    Helixi1707 – 171913Combined sources
    Helixi1725 – 17328Combined sources
    Helixi1734 – 17407Combined sources
    Beta strandi1749 – 175911Combined sources
    Turni1773 – 17764Combined sources
    Beta strandi1778 – 178811Combined sources
    Beta strandi1791 – 17955Combined sources
    Helixi1809 – 18113Combined sources
    Helixi1813 – 18153Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    4BY3X-ray1.73A1456-1846[»]
    4C6BX-ray1.66A1456-1846[»]
    4C6CX-ray1.45A1456-1846[»]
    4C6DX-ray1.30A1456-1846[»]
    4C6EX-ray1.26A1456-1846[»]
    4C6FX-ray1.26A1456-1846[»]
    4C6IX-ray1.35A1456-1846[»]
    4C6JX-ray1.30A1456-1846[»]
    4C6KX-ray1.48A1456-1846[»]
    4C6LX-ray1.55A1456-1846[»]
    4C6MX-ray1.62A1456-1846[»]
    4C6NX-ray1.90A1456-1846[»]
    4C6OX-ray1.65A1456-1846[»]
    4C6PX-ray1.52A1456-1846[»]
    4C6QX-ray1.66A1456-1846[»]
    ProteinModelPortaliP27708.
    SMRiP27708. Positions 2-359, 394-1823, 1923-2223.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini177 – 363187Glutamine amidotransferase type-1Add
    BLAST
    Domaini519 – 711193ATP-grasp 1Add
    BLAST
    Domaini1052 – 1243192ATP-grasp 2Add
    BLAST

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni2 – 365364GATase (Glutamine amidotransferase)Add
    BLAST
    Regioni366 – 39429LinkerAdd
    BLAST
    Regioni395 – 14551061CPSase (Carbamoyl-phosphate synthase)Add
    BLAST
    Regioni395 – 933539CPSase AAdd
    BLAST
    Regioni934 – 1455522CPSase BAdd
    BLAST
    Regioni1456 – 1788333DHOase (dihydroorotase)Add
    BLAST
    Regioni1789 – 1917129LinkerAdd
    BLAST
    Regioni1918 – 2225308ATCase (Aspartate transcarbamylase)Add
    BLAST

    Sequence similaritiesi

    In the central section; belongs to the DHOase family.Curated
    Contains 2 ATP-grasp domains.Curated

    Keywords - Domaini

    Repeat

    Phylogenomic databases

    eggNOGiCOG0458.
    GeneTreeiENSGT00390000015604.
    HOGENOMiHOG000234584.
    HOVERGENiHBG000279.
    InParanoidiP27708.
    KOiK11540.
    OrthoDBiEOG7M6D6F.
    PhylomeDBiP27708.
    TreeFamiTF105604.

    Family and domain databases

    Gene3Di1.10.1030.10. 1 hit.
    3.30.1490.20. 1 hit.
    3.30.470.20. 2 hits.
    3.40.50.1370. 2 hits.
    3.40.50.1380. 1 hit.
    3.40.50.20. 2 hits.
    3.40.50.880. 1 hit.
    3.50.30.20. 1 hit.
    HAMAPiMF_00001. Asp_carb_tr.
    MF_01209. CPSase_S_chain.
    InterProiIPR006132. Asp/Orn_carbamoyltranf_P-bd.
    IPR006130. Asp/Orn_carbamoylTrfase.
    IPR002082. Asp_carbamoyltransf.
    IPR006131. Asp_carbamoyltransf_Asp/Orn-bd.
    IPR011761. ATP-grasp.
    IPR013815. ATP_grasp_subdomain_1.
    IPR013816. ATP_grasp_subdomain_2.
    IPR006275. CarbamoylP_synth_lsu.
    IPR005481. CarbamoylP_synth_lsu_N.
    IPR005480. CarbamoylP_synth_lsu_oligo.
    IPR006274. CarbamoylP_synth_ssu.
    IPR002474. CarbamoylP_synth_ssu_N.
    IPR005479. CbamoylP_synth_lsu-like_ATP-bd.
    IPR005483. CbamoylP_synth_lsu_CPSase_dom.
    IPR029062. Class_I_gatase-like.
    IPR002195. Dihydroorotase_CS.
    IPR017926. GATASE.
    IPR011059. Metal-dep_hydrolase_composite.
    IPR011607. MGS-like_dom.
    IPR016185. PreATP-grasp_dom.
    [Graphical view]
    PfamiPF00289. CPSase_L_chain. 2 hits.
    PF02786. CPSase_L_D2. 2 hits.
    PF02787. CPSase_L_D3. 1 hit.
    PF00988. CPSase_sm_chain. 1 hit.
    PF00117. GATase. 1 hit.
    PF02142. MGS. 1 hit.
    PF00185. OTCace. 1 hit.
    PF02729. OTCace_N. 1 hit.
    [Graphical view]
    PRINTSiPR00100. AOTCASE.
    PR00101. ATCASE.
    PR00098. CPSASE.
    SMARTiSM01096. CPSase_L_D3. 1 hit.
    SM01097. CPSase_sm_chain. 1 hit.
    SM00851. MGS. 1 hit.
    [Graphical view]
    SUPFAMiSSF48108. SSF48108. 1 hit.
    SSF51338. SSF51338. 1 hit.
    SSF52021. SSF52021. 1 hit.
    SSF52317. SSF52317. 1 hit.
    SSF52335. SSF52335. 1 hit.
    SSF52440. SSF52440. 2 hits.
    SSF53671. SSF53671. 1 hit.
    TIGRFAMsiTIGR00670. asp_carb_tr. 1 hit.
    TIGR01369. CPSaseII_lrg. 1 hit.
    TIGR01368. CPSaseIIsmall. 1 hit.
    PROSITEiPS50975. ATP_GRASP. 2 hits.
    PS00097. CARBAMOYLTRANSFERASE. 1 hit.
    PS00866. CPSASE_1. 2 hits.
    PS00867. CPSASE_2. 2 hits.
    PS00482. DIHYDROOROTASE_1. 1 hit.
    PS00483. DIHYDROOROTASE_2. 1 hit.
    PS51273. GATASE_TYPE_1. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    P27708-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MAALVLEDGS VLRGQPFGAA VSTAGEVVFQ TGMVGYPEAL TDPSYKAQIL
    60 70 80 90 100
    VLTYPLIGNY GIPPDEMDEF GLCKWFESSG IHVAALVVGE CCPTPSHWSA
    110 120 130 140 150
    TRTLHEWLQQ HGIPGLQGVD TRELTKKLRE QGSLLGKLVQ NGTEPSSLPF
    160 170 180 190 200
    LDPNARPLVP EVSIKTPRVF NTGGAPRILA LDCGLKYNQI RCLCQRGAEV
    210 220 230 240 250
    TVVPWDHALD SQEYEGLFLS NGPGDPASYP SVVSTLSRVL SEPNPRPVFG
    260 270 280 290 300
    ICLGHQLLAL AIGAKTYKMR YGNRGHNQPC LLVGSGRCFL TSQNHGFAVE
    310 320 330 340 350
    TDSLPADWAP LFTNANDGSN EGIVHNSLPF FSVQFHPEHQ AGPSDMELLF
    360 370 380 390 400
    DIFLETVKEA TAGNPGGQTV RERLTERLCP PGIPTPGSGL PPPRKVLILG
    410 420 430 440 450
    SGGLSIGQAG EFDYSGSQAI KALKEENIQT LLINPNIATV QTSQGLADKV
    460 470 480 490 500
    YFLPITPHYV TQVIRNERPD GVLLTFGGQT ALNCGVELTK AGVLARYGVR
    510 520 530 540 550
    VLGTPVETIE LTEDRRAFAA RMAEIGEHVA PSEAANSLEQ AQAAAERLGY
    560 570 580 590 600
    PVLVRAAFAL GGLGSGFASN REELSALVAP AFAHTSQVLV DKSLKGWKEI
    610 620 630 640 650
    EYEVVRDAYG NCVTVCNMEN LDPLGIHTGE SIVVAPSQTL NDREYQLLRQ
    660 670 680 690 700
    TAIKVTQHLG IVGECNVQYA LNPESEQYYI IEVNARLSRS SALASKATGY
    710 720 730 740 750
    PLAYVAAKLA LGIPLPELRN SVTGGTAAFE PSVDYCVVKI PRWDLSKFLR
    760 770 780 790 800
    VSTKIGSCMK SVGEVMGIGR SFEEAFQKAL RMVDENCVGF DHTVKPVSDM
    810 820 830 840 850
    ELETPTDKRI FVVAAALWAG YSVDRLYELT RIDRWFLHRM KRIIAHAQLL
    860 870 880 890 900
    EQHRGQPLPP DLLQQAKCLG FSDKQIALAV LSTELAVRKL RQELGICPAV
    910 920 930 940 950
    KQIDTVAAEW PAQTNYLYLT YWGTTHDLTF RTPHVLVLGS GVYRIGSSVE
    960 970 980 990 1000
    FDWCAVGCIQ QLRKMGYKTI MVNYNPETVS TDYDMCDRLY FDEISFEVVM
    1010 1020 1030 1040 1050
    DIYELENPEG VILSMGGQLP NNMAMALHRQ QCRVLGTSPE AIDSAENRFK
    1060 1070 1080 1090 1100
    FSRLLDTIGI SQPQWRELSD LESARQFCQT VGYPCVVRPS YVLSGAAMNV
    1110 1120 1130 1140 1150
    AYTDGDLERF LSSAAAVSKE HPVVISKFIQ EAKEIDVDAV ASDGVVAAIA
    1160 1170 1180 1190 1200
    ISEHVENAGV HSGDATLVTP PQDITAKTLE RIKAIVHAVG QELQVTGPFN
    1210 1220 1230 1240 1250
    LQLIAKDDQL KVIECNVRVS RSFPFVSKTL GVDLVALATR VIMGEEVEPV
    1260 1270 1280 1290 1300
    GLMTGSGVVG VKVPQFSFSR LAGADVVLGV EMTSTGEVAG FGESRCEAYL
    1310 1320 1330 1340 1350
    KAMLSTGFKI PKKNILLTIG SYKNKSELLP TVRLLESLGY SLYASLGTAD
    1360 1370 1380 1390 1400
    FYTEHGVKVT AVDWHFEEAV DGECPPQRSI LEQLAEKNFE LVINLSMRGA
    1410 1420 1430 1440 1450
    GGRRLSSFVT KGYRTRRLAA DFSVPLIIDI KCTKLFVEAL GQIGPAPPLK
    1460 1470 1480 1490 1500
    VHVDCMTSQK LVRLPGLIDV HVHLREPGGT HKEDFASGTA AALAGGITMV
    1510 1520 1530 1540 1550
    CAMPNTRPPI IDAPALALAQ KLAEAGARCD FALFLGASSE NAGTLGTVAG
    1560 1570 1580 1590 1600
    SAAGLKLYLN ETFSELRLDS VVQWMEHFET WPSHLPIVAH AEQQTVAAVL
    1610 1620 1630 1640 1650
    MVAQLTQRSV HICHVARKEE ILLIKAAKAR GLPVTCEVAP HHLFLSHDDL
    1660 1670 1680 1690 1700
    ERLGPGKGEV RPELGSRQDV EALWENMAVI DCFASDHAPH TLEEKCGSRP
    1710 1720 1730 1740 1750
    PPGFPGLETM LPLLLTAVSE GRLSLDDLLQ RLHHNPRRIF HLPPQEDTYV
    1760 1770 1780 1790 1800
    EVDLEHEWTI PSHMPFSKAH WTPFEGQKVK GTVRRVVLRG EVAYIDGQVL
    1810 1820 1830 1840 1850
    VPPGYGQDVR KWPQGAVPQL PPSAPATSEM TTTPERPRRG IPGLPDGRFH
    1860 1870 1880 1890 1900
    LPPRIHRASD PGLPAEEPKE KSSRKVAEPE LMGTPDGTCY PPPPVPRQAS
    1910 1920 1930 1940 1950
    PQNLGTPGLL HPQTSPLLHS LVGQHILSVQ QFTKDQMSHL FNVAHTLRMM
    1960 1970 1980 1990 2000
    VQKERSLDIL KGKVMASMFY EVSTRTSSSF AAAMARLGGA VLSFSEATSS
    2010 2020 2030 2040 2050
    VQKGESLADS VQTMSCYADV VVLRHPQPGA VELAAKHCRR PVINAGDGVG
    2060 2070 2080 2090 2100
    EHPTQALLDI FTIREELGTV NGMTITMVGD LKHGRTVHSL ACLLTQYRVS
    2110 2120 2130 2140 2150
    LRYVAPPSLR MPPTVRAFVA SRGTKQEEFE SIEEALPDTD VLYMTRIQKE
    2160 2170 2180 2190 2200
    RFGSTQEYEA CFGQFILTPH IMTRAKKKMV VMHPMPRVNE ISVEVDSDPR
    2210 2220
    AAYFRQAENG MYIRMALLAT VLGRF
    Length:2,225
    Mass (Da):242,984
    Last modified:July 19, 2004 - v3
    Checksum:i2AB8E8413E825A8F
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti505 – 5051P → T in BAA11423 (PubMed:8619816).Curated
    Sequence conflicti535 – 5351A → G in BAA11423 (PubMed:8619816).Curated
    Sequence conflicti560 – 5601L → V in BAA11423 (PubMed:8619816).Curated
    Sequence conflicti1103 – 11031T → A in BAA11423 (PubMed:8619816).Curated
    Sequence conflicti1513 – 15131A → G in BAA11423 (PubMed:8619816).Curated
    Sequence conflicti1676 – 16761N → D in BAA11423 (PubMed:8619816).Curated

    Natural variant

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Natural varianti177 – 1771R → Q in a colorectal cancer sample; somatic mutation. 1 Publication
    VAR_035897
    Natural varianti735 – 7351Y → C in a colorectal cancer sample; somatic mutation. 1 Publication
    VAR_035898

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D78586 mRNA. Translation: BAA11423.1.
    CH471053 Genomic DNA. Translation: EAX00612.1.
    CH471053 Genomic DNA. Translation: EAX00613.1.
    CH471053 Genomic DNA. Translation: EAX00614.1.
    BC014178 mRNA. Translation: AAH14178.2.
    BC065510 mRNA. Translation: AAH65510.1.
    M38561 Genomic DNA. Translation: AAA51907.1.
    CCDSiCCDS1742.1.
    PIRiA36240.
    RefSeqiNP_001293008.1. NM_001306079.1.
    NP_004332.2. NM_004341.4.
    UniGeneiHs.377010.

    Genome annotation databases

    EnsembliENST00000264705; ENSP00000264705; ENSG00000084774.
    GeneIDi790.
    KEGGihsa:790.
    UCSCiuc002rji.3. human.

    Keywords - Coding sequence diversityi

    Polymorphism

    Cross-referencesi

    Web resourcesi

    Wikipedia

    Aspartate carbamoyltransferase entry

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D78586 mRNA. Translation: BAA11423.1.
    CH471053 Genomic DNA. Translation: EAX00612.1.
    CH471053 Genomic DNA. Translation: EAX00613.1.
    CH471053 Genomic DNA. Translation: EAX00614.1.
    BC014178 mRNA. Translation: AAH14178.2.
    BC065510 mRNA. Translation: AAH65510.1.
    M38561 Genomic DNA. Translation: AAA51907.1.
    CCDSiCCDS1742.1.
    PIRiA36240.
    RefSeqiNP_001293008.1. NM_001306079.1.
    NP_004332.2. NM_004341.4.
    UniGeneiHs.377010.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    4BY3X-ray1.73A1456-1846[»]
    4C6BX-ray1.66A1456-1846[»]
    4C6CX-ray1.45A1456-1846[»]
    4C6DX-ray1.30A1456-1846[»]
    4C6EX-ray1.26A1456-1846[»]
    4C6FX-ray1.26A1456-1846[»]
    4C6IX-ray1.35A1456-1846[»]
    4C6JX-ray1.30A1456-1846[»]
    4C6KX-ray1.48A1456-1846[»]
    4C6LX-ray1.55A1456-1846[»]
    4C6MX-ray1.62A1456-1846[»]
    4C6NX-ray1.90A1456-1846[»]
    4C6OX-ray1.65A1456-1846[»]
    4C6PX-ray1.52A1456-1846[»]
    4C6QX-ray1.66A1456-1846[»]
    ProteinModelPortaliP27708.
    SMRiP27708. Positions 2-359, 394-1823, 1923-2223.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    BioGridi107243. 44 interactions.
    DIPiDIP-39484N.
    IntActiP27708. 25 interactions.
    MINTiMINT-5000537.
    STRINGi9606.ENSP00000264705.

    Chemistry

    BindingDBiP27708.
    ChEMBLiCHEMBL3093.
    DrugBankiDB00128. L-Aspartic Acid.
    DB00130. L-Glutamine.

    Protein family/group databases

    MEROPSiM38.972.

    PTM databases

    PhosphoSiteiP27708.

    Polymorphism and mutation databases

    BioMutaiCAD.
    DMDMi50403731.

    Proteomic databases

    MaxQBiP27708.
    PaxDbiP27708.
    PeptideAtlasiP27708.
    PRIDEiP27708.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsembliENST00000264705; ENSP00000264705; ENSG00000084774.
    GeneIDi790.
    KEGGihsa:790.
    UCSCiuc002rji.3. human.

    Organism-specific databases

    CTDi790.
    GeneCardsiGC02P027440.
    HGNCiHGNC:1424. CAD.
    HPAiCAB007781.
    HPA050746.
    HPA057266.
    MIMi114010. gene.
    neXtProtiNX_P27708.
    PharmGKBiPA26023.
    GenAtlasiSearch...

    Phylogenomic databases

    eggNOGiCOG0458.
    GeneTreeiENSGT00390000015604.
    HOGENOMiHOG000234584.
    HOVERGENiHBG000279.
    InParanoidiP27708.
    KOiK11540.
    OrthoDBiEOG7M6D6F.
    PhylomeDBiP27708.
    TreeFamiTF105604.

    Enzyme and pathway databases

    UniPathwayiUPA00070; UER00115.
    UPA00070; UER00116.
    UPA00070; UER00117.
    BioCyciMetaCyc:ENSG00000084774-MONOMER.
    BRENDAi3.5.2.3. 2681.
    ReactomeiREACT_21376. Pyrimidine biosynthesis.

    Miscellaneous databases

    ChiTaRSiCAD. human.
    GenomeRNAii790.
    NextBioi3214.
    PMAP-CutDBP27708.
    PROiP27708.
    SOURCEiSearch...

    Gene expression databases

    BgeeiP27708.
    CleanExiHS_CAD.
    ExpressionAtlasiP27708. baseline and differential.
    GenevisibleiP27708. HS.

    Family and domain databases

    Gene3Di1.10.1030.10. 1 hit.
    3.30.1490.20. 1 hit.
    3.30.470.20. 2 hits.
    3.40.50.1370. 2 hits.
    3.40.50.1380. 1 hit.
    3.40.50.20. 2 hits.
    3.40.50.880. 1 hit.
    3.50.30.20. 1 hit.
    HAMAPiMF_00001. Asp_carb_tr.
    MF_01209. CPSase_S_chain.
    InterProiIPR006132. Asp/Orn_carbamoyltranf_P-bd.
    IPR006130. Asp/Orn_carbamoylTrfase.
    IPR002082. Asp_carbamoyltransf.
    IPR006131. Asp_carbamoyltransf_Asp/Orn-bd.
    IPR011761. ATP-grasp.
    IPR013815. ATP_grasp_subdomain_1.
    IPR013816. ATP_grasp_subdomain_2.
    IPR006275. CarbamoylP_synth_lsu.
    IPR005481. CarbamoylP_synth_lsu_N.
    IPR005480. CarbamoylP_synth_lsu_oligo.
    IPR006274. CarbamoylP_synth_ssu.
    IPR002474. CarbamoylP_synth_ssu_N.
    IPR005479. CbamoylP_synth_lsu-like_ATP-bd.
    IPR005483. CbamoylP_synth_lsu_CPSase_dom.
    IPR029062. Class_I_gatase-like.
    IPR002195. Dihydroorotase_CS.
    IPR017926. GATASE.
    IPR011059. Metal-dep_hydrolase_composite.
    IPR011607. MGS-like_dom.
    IPR016185. PreATP-grasp_dom.
    [Graphical view]
    PfamiPF00289. CPSase_L_chain. 2 hits.
    PF02786. CPSase_L_D2. 2 hits.
    PF02787. CPSase_L_D3. 1 hit.
    PF00988. CPSase_sm_chain. 1 hit.
    PF00117. GATase. 1 hit.
    PF02142. MGS. 1 hit.
    PF00185. OTCace. 1 hit.
    PF02729. OTCace_N. 1 hit.
    [Graphical view]
    PRINTSiPR00100. AOTCASE.
    PR00101. ATCASE.
    PR00098. CPSASE.
    SMARTiSM01096. CPSase_L_D3. 1 hit.
    SM01097. CPSase_sm_chain. 1 hit.
    SM00851. MGS. 1 hit.
    [Graphical view]
    SUPFAMiSSF48108. SSF48108. 1 hit.
    SSF51338. SSF51338. 1 hit.
    SSF52021. SSF52021. 1 hit.
    SSF52317. SSF52317. 1 hit.
    SSF52335. SSF52335. 1 hit.
    SSF52440. SSF52440. 2 hits.
    SSF53671. SSF53671. 1 hit.
    TIGRFAMsiTIGR00670. asp_carb_tr. 1 hit.
    TIGR01369. CPSaseII_lrg. 1 hit.
    TIGR01368. CPSaseIIsmall. 1 hit.
    PROSITEiPS50975. ATP_GRASP. 2 hits.
    PS00097. CARBAMOYLTRANSFERASE. 1 hit.
    PS00866. CPSASE_1. 2 hits.
    PS00867. CPSASE_2. 2 hits.
    PS00482. DIHYDROOROTASE_1. 1 hit.
    PS00483. DIHYDROOROTASE_2. 1 hit.
    PS51273. GATASE_TYPE_1. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    1. "Molecular cloning of a human cDNA encoding a trifunctional enzyme of carbamoyl-phosphate synthetase-aspartate transcarbamoylase-dihydroorotase in de Novo pyrimidine synthesis."
      Iwahana H., Fujimura M., Ii S., Kondo M., Moritani M., Takahashi Y., Yamaoka T., Yoshimoto K., Itakura M.
      Biochem. Biophys. Res. Commun. 219:249-255(1996) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [MRNA].
      Tissue: Fetal lung fibroblast.
    2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    3. "The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC)."
      The MGC Project Team
      Genome Res. 14:2121-2127(2004) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
      Tissue: Eye.
    4. Cited for: PROTEIN SEQUENCE OF 2-13; 157-165; 169-177; 359-371; 501-516; 548-555; 599-606; 697-719; 761-778; 843-854; 932-944; 1034-1048; 1067-1075; 1110-1127; 1229-1240; 1263-1270; 1313-1323; 1326-1333; 1658-1667; 1723-1731; 1840-1848; 1976-1986; 2025-2036; 2103-2110; 2179-2187 AND 2215-2225, CLEAVAGE OF INITIATOR METHIONINE, ACETYLATION AT ALA-2, IDENTIFICATION BY MASS SPECTROMETRY.
      Tissue: Colon adenocarcinoma and Hepatoma.
    5. "Organization and nucleotide sequence of the 3' end of the human CAD gene."
      Davidson J.N., Rao G.N., Niswander L., Andreano C., Tamer C., Chen K.C.
      DNA Cell Biol. 9:667-676(1990) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1752-2225.
    6. "Function of conserved histidine residues in mammalian dihydroorotase."
      Zimmermann B.H., Kemling N.M., Evans D.R.
      Biochemistry 34:7038-7046(1995) [PubMed] [Europe PMC] [Abstract]
      Cited for: MUTAGENESIS OF HIS-1471; HIS-1473; ASP-1512; HIS-1590; HIS-1642 AND HIS-1690, COFACTOR, ZINC-BINDING SITES.
    7. "Growth-dependent regulation of mammalian pyrimidine biosynthesis by the protein kinase A and MAPK signaling cascades."
      Sigoillot F.D., Evans D.R., Guy H.I.
      J. Biol. Chem. 277:15745-15751(2002) [PubMed] [Europe PMC] [Abstract]
      Cited for: ENZYME REGULATION.
    8. "Nuclear localization and mitogen-activated protein kinase phosphorylation of the multifunctional protein CAD."
      Sigoillot F.D., Kotsis D.H., Serre V., Sigoillot S.M., Evans D.R., Guy H.I.
      J. Biol. Chem. 280:25611-25620(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: SUBCELLULAR LOCATION.
    9. "Transcriptional repression of human cad gene by hypoxia inducible factor-1alpha."
      Chen K.F., Lai Y.Y., Sun H.S., Tsai S.J.
      Nucleic Acids Res. 33:5190-5198(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: INDUCTION.
    10. "Global, in vivo, and site-specific phosphorylation dynamics in signaling networks."
      Olsen J.V., Blagoev B., Gnad F., Macek B., Kumar C., Mortensen P., Mann M.
      Cell 127:635-648(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1859, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Cervix carcinoma.
    11. "A probability-based approach for high-throughput protein phosphorylation analysis and site localization."
      Beausoleil S.A., Villen J., Gerber S.A., Rush J., Gygi S.P.
      Nat. Biotechnol. 24:1285-1292(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1859, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Cervix carcinoma.
    12. "Protein kinase C modulates the up-regulation of the pyrimidine biosynthetic complex, CAD, by MAP kinase."
      Sigoillot F.D., Kotsis D.H., Masko E.M., Bame M., Evans D.R., Evans H.I.
      Front. Biosci. 12:3892-3898(2007) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION AT THR-456; SER-1406; SER-1859 AND SER-1873, MUTAGENESIS OF SER-1873.
    13. "Kinase-selective enrichment enables quantitative phosphoproteomics of the kinome across the cell cycle."
      Daub H., Olsen J.V., Bairlein M., Gnad F., Oppermann F.S., Korner R., Greff Z., Keri G., Stemmann O., Mann M.
      Mol. Cell 31:438-448(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1859 AND THR-1884, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Cervix carcinoma.
    14. Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT THR-1884, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Cervix carcinoma.
    15. "Lys-N and trypsin cover complementary parts of the phosphoproteome in a refined SCX-based approach."
      Gauci S., Helbig A.O., Slijper M., Krijgsveld J., Heck A.J., Mohammed S.
      Anal. Chem. 81:4493-4501(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS], CLEAVAGE OF INITIATOR METHIONINE [LARGE SCALE ANALYSIS].
    16. "Quantitative phosphoproteomic analysis of T cell receptor signaling reveals system-wide modulation of protein-protein interactions."
      Mayya V., Lundgren D.H., Hwang S.-I., Rezaul K., Wu L., Eng J.K., Rodionov V., Han D.K.
      Sci. Signal. 2:RA46-RA46(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Leukemic T-cell.
    17. "Lysine acetylation targets protein complexes and co-regulates major cellular functions."
      Choudhary C., Kumar C., Gnad F., Nielsen M.L., Rehman M., Walther T.C., Olsen J.V., Mann M.
      Science 325:834-840(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: ACETYLATION [LARGE SCALE ANALYSIS] AT LYS-747 AND LYS-1411, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
    18. "Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis."
      Olsen J.V., Vermeulen M., Santamaria A., Kumar C., Miller M.L., Jensen L.J., Gnad F., Cox J., Jensen T.S., Nigg E.A., Brunak S., Mann M.
      Sci. Signal. 3:RA3-RA3(2010) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1859 AND SER-1900, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Cervix carcinoma.
    19. Cited for: IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
    20. "System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation."
      Rigbolt K.T., Prokhorova T.A., Akimov V., Henningsen J., Johansen P.T., Kratchmarova I., Kassem M., Mann M., Olsen J.V., Blagoev B.
      Sci. Signal. 4:RS3-RS3(2011) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1406 AND SER-1859, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
    21. "Comparative large-scale characterisation of plant vs. mammal proteins reveals similar and idiosyncratic N-alpha acetylation features."
      Bienvenut W.V., Sumpton D., Martinez A., Lilla S., Espagne C., Meinnel T., Giglione C.
      Mol. Cell. Proteomics 11:M111.015131-M111.015131(2012) [PubMed] [Europe PMC] [Abstract]
      Cited for: ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS], CLEAVAGE OF INITIATOR METHIONINE [LARGE SCALE ANALYSIS].
    22. Cited for: ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS], CLEAVAGE OF INITIATOR METHIONINE [LARGE SCALE ANALYSIS].
    23. "Quantitative phosphoproteomics reveal mTORC1 activates de novo pyrimidine synthesis."
      Robitaille A.M., Christen S., Shimobayashi M., Cornu M., Fava L.L., Moes S., Prescianotto-Baschong C., Sauer U., Jenoe P., Hall M.N.
      Science 339:1320-1323(2013) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION AT SER-1859.
    24. "Stimulation of de novo pyrimidine synthesis by growth signaling through mTOR and S6K1."
      Ben-Sahra I., Howell J.J., Asara J.M., Manning B.D.
      Science 339:1323-1328(2013) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION AT SER-1859 AND SER-1900.
    25. "An enzyme assisted RP-RPLC approach for in-depth analysis of human liver phosphoproteome."
      Bian Y., Song C., Cheng K., Dong M., Wang F., Huang J., Sun D., Wang L., Ye M., Zou H.
      J. Proteomics 96:253-262(2014) [PubMed] [Europe PMC] [Abstract]
      Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-1859, IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
      Tissue: Liver.
    26. "Structure, functional characterization, and evolution of the dihydroorotase domain of human CAD."
      Grande-Garcia A., Lallous N., Diaz-Tejada C., Ramon-Maiques S.
      Structure 22:185-198(2014) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS) OF 1456-1846 IN COMPLEX WITH DIHYDROOROTATE; N-CARBAMOYL-L-ASPARTIC ACID AND ZINC, COFACTOR, CATALYTIC ACTIVITY, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF HIS-1471; THR-1562; PHE-1563; HIS-1590; CYS-1613; HIS-1614; GLU-1637 AND ASP-1686, CARBAMYLATION AT LYS-1556.
    27. Cited for: VARIANTS [LARGE SCALE ANALYSIS] GLN-177 AND CYS-735.

    Entry informationi

    Entry nameiPYR1_HUMAN
    AccessioniPrimary (citable) accession number: P27708
    Secondary accession number(s): D6W552, Q6P0Q0, Q96CK3
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: August 1, 1992
    Last sequence update: July 19, 2004
    Last modified: July 22, 2015
    This is version 180 of the entry and version 3 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programChordata Protein Annotation Program
    DisclaimerAny medical or genetic information present in this entry is provided for research, educational and informational purposes only. It is not in any way intended to be used as a substitute for professional medical advice, diagnosis, treatment or care.

    Miscellaneousi

    Miscellaneous

    GATase (glutamine amidotransferase) and CPSase (carbamoyl phosphate synthase) form together the glutamine-dependent CPSase (GD-CPSase) (EC 6.3.5.5).

    Keywords - Technical termi

    3D-structure, Allosteric enzyme, Complete proteome, Direct protein sequencing, Multifunctional enzyme, Reference proteome

    Documents

    1. Human chromosome 2
      Human chromosome 2: entries, gene names and cross-references to MIM
    2. Human entries with polymorphisms or disease mutations
      List of human entries with polymorphisms or disease mutations
    3. Human polymorphisms and disease mutations
      Index of human polymorphisms and disease mutations
    4. MIM cross-references
      Online Mendelian Inheritance in Man (MIM) cross-references in UniProtKB/Swiss-Prot
    5. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    6. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    7. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.