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- EMDB-15977: IFTB1 subcomplex of anterograde intraflagellar transport trains (... -

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Basic information

Entry
Database: EMDB / ID: EMD-15977
TitleIFTB1 subcomplex of anterograde intraflagellar transport trains (Chlamydomonas reinhardtii)
Map data
Sample
  • Complex: Two IFTB repeats from anterograde intraflagellar transport trains within native Chlamydomonas reinhardtii cilia
    • Protein or peptide: x 13 types
KeywordsCilia / IFT / Intraflagellar / transport / PROTEIN TRANSPORT
Function / homology
Function and homology information


intraciliary transport particle B binding / intraciliary transport involved in cilium assembly / intraciliary transport particle B / outer dynein arm assembly / cilium-dependent cell motility / intraciliary transport / regulation of cilium assembly / : / axoneme assembly / axonemal microtubule ...intraciliary transport particle B binding / intraciliary transport involved in cilium assembly / intraciliary transport particle B / outer dynein arm assembly / cilium-dependent cell motility / intraciliary transport / regulation of cilium assembly / : / axoneme assembly / axonemal microtubule / cilium organization / non-motile cilium assembly / non-motile cilium / motile cilium / ciliary membrane / ciliary base / post-transcriptional regulation of gene expression / beta-tubulin binding / axoneme / kinesin binding / cilium assembly / regulation of microtubule cytoskeleton organization / centriole / tubulin binding / ciliary basal body / regulation of protein stability / cilium / protein transport / apical part of cell / microtubule binding / cytoskeleton / protein stabilization / cytoplasm
Similarity search - Function
Tetratricopeptide repeat protein 26 / Clusterin-associated protein-1 / Intra-flagellar transport protein 57 / Intraflagellar transport protein 20 / Clusterin-associated protein-1 / Intra-flagellar transport protein 57 / Intraflagellar transport complex B, subunit 20 / Intraflagellar transport complex B protein 46 / Intraflagellar transport protein 74 / Tetratricopeptide repeat protein 30 ...Tetratricopeptide repeat protein 26 / Clusterin-associated protein-1 / Intra-flagellar transport protein 57 / Intraflagellar transport protein 20 / Clusterin-associated protein-1 / Intra-flagellar transport protein 57 / Intraflagellar transport complex B, subunit 20 / Intraflagellar transport complex B protein 46 / Intraflagellar transport protein 74 / Tetratricopeptide repeat protein 30 / : / Intraflagellar transport complex B protein 46 C terminal / Intraflagellar transport protein 52, C-terminal domain / ABC-type uncharacterised transport system / Intraflagellar transport protein 81 / IFT81, calponin homology domain / IFT81, N-terminal domain superfamily / ABC-type uncharacterized transport system / Intraflagellar transport 81 calponin homology domain / Intraflagellar transport protein 52 homolog / TRAF3-interacting protein 1, N-terminal / TRAF3-interacting protein 1, C-terminal domain / TRAF3-interacting protein 1, N-terminal domain superfamily / Microtubule-binding protein MIP-T3 CH-like domain / Microtubule-binding protein MIP-T3 C-terminal region / TRAF3-interacting protein 1 / Tetratricopeptide repeat / Quinoprotein amine dehydrogenase, beta chain-like / Tetratricopeptide repeat / Tetratricopeptide repeat / IQ motif, EF-hand binding site / Class I glutamine amidotransferase-like / TPR repeat profile. / Tetratricopeptide repeats / Tetratricopeptide repeat / Tetratricopeptide-like helical domain superfamily / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD domain, G-beta repeat / WD40 repeats / WD40 repeat / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
Clusterin-associated protein 1 / Intraflagellar transport protein 46 / Intraflagellar transport protein 80 / Uncharacterized protein / Intraflagellar transport protein 56 / Uncharacterized protein / Intraflagellar transport protein 88 / Intraflagellar transport protein 57 / Intraflagellar transport protein 172 / Intraflagellar transport protein 81 ...Clusterin-associated protein 1 / Intraflagellar transport protein 46 / Intraflagellar transport protein 80 / Uncharacterized protein / Intraflagellar transport protein 56 / Uncharacterized protein / Intraflagellar transport protein 88 / Intraflagellar transport protein 57 / Intraflagellar transport protein 172 / Intraflagellar transport protein 81 / Intraflagellar transport protein 74 / Intraflagellar transport particle protein IFT20 / Osm-6-like protein
Similarity search - Component
Biological speciesChlamydomonas reinhardtii (plant)
Methodsubtomogram averaging / cryo EM / Resolution: 9.9 Å
AuthorsLacey SE / Foster HE / Pigino G
Funding supportEuropean Union, Italy, 2 items
OrganizationGrant numberCountry
European Research Council (ERC)European Union
Other government Italy
CitationJournal: Nat Struct Mol Biol / Year: 2023
Title: The molecular structure of IFT-A and IFT-B in anterograde intraflagellar transport trains.
Authors: Samuel E Lacey / Helen E Foster / Gaia Pigino /
Abstract: Anterograde intraflagellar transport (IFT) trains are essential for cilia assembly and maintenance. These trains are formed of 22 IFT-A and IFT-B proteins that link structural and signaling cargos to ...Anterograde intraflagellar transport (IFT) trains are essential for cilia assembly and maintenance. These trains are formed of 22 IFT-A and IFT-B proteins that link structural and signaling cargos to microtubule motors for import into cilia. It remains unknown how the IFT-A/-B proteins are arranged into complexes and how these complexes polymerize into functional trains. Here we use in situ cryo-electron tomography of Chlamydomonas reinhardtii cilia and AlphaFold2 protein structure predictions to generate a molecular model of the entire anterograde train. We show how the conformations of both IFT-A and IFT-B are dependent on lateral interactions with neighboring repeats, suggesting that polymerization is required to cooperatively stabilize the complexes. Following three-dimensional classification, we reveal how IFT-B extends two flexible tethers to maintain a connection with IFT-A that can withstand the mechanical stresses present in actively beating cilia. Overall, our findings provide a framework for understanding the fundamental processes that govern cilia assembly.
History
DepositionOct 18, 2022-
Header (metadata) releaseJan 11, 2023-
Map releaseJan 11, 2023-
UpdateJul 26, 2023-
Current statusJul 26, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_15977.png

Downloads & links

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Map

FileDownload / File: emd_15977.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 3.03 Å
Density
Contour LevelBy AUTHOR: 0.466
Minimum - Maximum-0.38829696 - 1.3724803
Average (Standard dev.)0.0042394185 (±0.044678286)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 775.68 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Two IFTB repeats from anterograde intraflagellar transport trains...

EntireName: Two IFTB repeats from anterograde intraflagellar transport trains within native Chlamydomonas reinhardtii cilia
Components
  • Complex: Two IFTB repeats from anterograde intraflagellar transport trains within native Chlamydomonas reinhardtii cilia
    • Protein or peptide: IFT88
    • Protein or peptide: Osm-6-like protein
    • Protein or peptide: IFT70
    • Protein or peptide: Intraflagellar transport protein 46
    • Protein or peptide: Intraflagellar transport protein 56
    • Protein or peptide: Intraflagellar transport protein 81
    • Protein or peptide: Intraflagellar transport protein 74
    • Protein or peptide: Intraflagellar transport protein 80
    • Protein or peptide: Clusterin-associated protein 1
    • Protein or peptide: Intraflagellar transport protein 57
    • Protein or peptide: Intraflagellar transport particle protein IFT20
    • Protein or peptide: IFT54
    • Protein or peptide: Intraflagellar transport protein 172

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Supramolecule #1: Two IFTB repeats from anterograde intraflagellar transport trains...

SupramoleculeName: Two IFTB repeats from anterograde intraflagellar transport trains within native Chlamydomonas reinhardtii cilia
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Chlamydomonas reinhardtii (plant)

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Macromolecule #1: IFT88

MacromoleculeName: IFT88 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 86.418125 KDa
SequenceString: MSYGGTEEDD LYGGYDEQSN PLAGSGGAAF KALGADGAPP GTAMMGPPGT AMKSFVPGTA MRGGTAMQQD PSLARPMTSN RGAGFTSAP NKKFDPLNRS MGSTLGSSGG GAMLVARKGD TSPEEQARGM EKTVHELLEK SAADAAKNDI NSALENAMEA K KNERKLCR ...String:
MSYGGTEEDD LYGGYDEQSN PLAGSGGAAF KALGADGAPP GTAMMGPPGT AMKSFVPGTA MRGGTAMQQD PSLARPMTSN RGAGFTSAP NKKFDPLNRS MGSTLGSSGG GAMLVARKGD TSPEEQARGM EKTVHELLEK SAADAAKNDI NSALENAMEA K KNERKLCR FREQNNMADQ INLELMYAVD FNLAHMYHMN KNYSEALNLY TAIVRNKNFP QSGWLRVNMG NIHFEQKKYP SA IKMYRMA LDQISATAKE VRFKIMRNIG LSFVRMGQYP DALQSFATVM DNVPDHQTGY NLVMCNYALS DREGMKNAFI KLL KVSPPS EMDDDDDDDP MGDDDMQVMT MDDGLKDEMR KRNTIITRLI VKAAQLISEK VDRANGFEGG FMWCCEQLRD AGYT KLANE VELAKATRFM GQKQFDKAVG VFKDFEKKEP RVKARAATNL AFLYFLEGET DQADKYSEMA LKSDRYNARA YVNKG CVLV ERGDLEGARS LFNEAAGIDP YCVEAIYNLG LVSQRLNELP YALAAFKKLH NMVPDNVEVI HQIATTYDMM GDFKNA VKW FELLTSLVSN DPGVLARLGA IHARFDDEAK ALHYYQESHR VYPVNMDVIS WLGAYHVKSE VYEKAMPFFD LASKIQP QE VKWALMVASC YRRTNNLPAA LGKYKQIHTQ HPDNVECLRY LVHLCSELGR RAEAAEYMTK LKKAEKAAVP EATTAAAP A AAAAGSGMGG MGGLDDDIGS SAVSAQNRGK KMLVKEHMGG GGGKDNDDWG NEQLGDDLLP M

UniProtKB: Intraflagellar transport protein 88

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Macromolecule #2: Osm-6-like protein

MacromoleculeName: Osm-6-like protein / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 50.453184 KDa
SequenceString: MEEPGAEEVR ILFSTAKGES HTHKAGFKQL FRRLRSTYRP DKVDKDDFTL DTLRSAHILV LGGPKEKFTA PEVDMLKKFV KNGGSILIL MSEGGEEKAG TNINYFLEQF GMSVNNDAVV RTTHYKYLHP KEVLISDGIL NRAVITGAGK SLNSNDDDEF R VSRGPQAF ...String:
MEEPGAEEVR ILFSTAKGES HTHKAGFKQL FRRLRSTYRP DKVDKDDFTL DTLRSAHILV LGGPKEKFTA PEVDMLKKFV KNGGSILIL MSEGGEEKAG TNINYFLEQF GMSVNNDAVV RTTHYKYLHP KEVLISDGIL NRAVITGAGK SLNSNDDDEF R VSRGPQAF DGTGLEYVFP FGATLSVQKP AVPVLSSGKI AYPMNRPVGA VWAQPGYGRI AVLGSCAMFD DKWLDKEENS KI MDFFFKF LKPHSKIQLN DIDAEEPDVS DLKLLPDTAS LADKLKGCLQ EIDDVPRDWT SLFDDSLFKF DTGLIPEAVS LYE KLGVKK GQLNLIPPSF ETPLPPLQPA VFPPTIREPP PPALELFDLD ESFASETNRL ASLTNKCHGE EDLEYYIMEA GHIL GLKLQ ENANAKHVLS EVFRRIAQYK MGSLGLGQTL DSMGQTLPAA NQFGDQFEL

UniProtKB: Osm-6-like protein

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Macromolecule #3: IFT70

MacromoleculeName: IFT70 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 74.248609 KDa
SequenceString: MAFFQQPARP IAEGQYTQTI YTLIKEQKFA EAIQHLQYQL QNVPESRAAL SLLGYCYYYT GQYDMASQMY EQLVTLYPSN EDYKLYYAQ SLYKGGMYPE ASKAVVKVEG HQKAVTTLLV ACSYEQDDLT GCRRQLDKCA PEDPDTMVNT GCIMFKEGKF E AARQKFND ...String:
MAFFQQPARP IAEGQYTQTI YTLIKEQKFA EAIQHLQYQL QNVPESRAAL SLLGYCYYYT GQYDMASQMY EQLVTLYPSN EDYKLYYAQ SLYKGGMYPE ASKAVVKVEG HQKAVTTLLV ACSYEQDDLT GCRRQLDKCA PEDPDTMVNT GCIMFKEGKF E AARQKFND AVQALGYQPE LLYNIALCYY KTKQFGPALK HLAEIIEKAV REHPELSVGS NTDGMEVRSV GNSQTLKETA LI EAFNLKA AIEYTMKNVE AAKEALTDMP PRAEEELDPV TLHNSALINM DSDPTGGFKK LNFLLQSPPF PPETFANLLL LYC KPSHGF YDLAADVLAE NPQYAGKLLS PDLYDYLQAA IGRYKSPEEA FRRFDELATR HVEQLRRLTK QIQDARIARD NDAI KRAIN EYDEALEAYI PGLMAMASIY WDMELYSNVE KIFRQSAEFC SEHEVWKLNV AHTFFMQDNH YKEAIRYYEP VVKKN ADNL LGVTAIVLAN LCVSYIMTSQ NEEAEELMRK VEKEEERSSM QDPDKPCFHL CIINLVIGTL YCAKGNYEFG VSRIIK SLE PYDKKLETDT WYYAKRCFLA LIENLAKHMI VLKDSSFTEI MAFLNEAEKH GKDIRVVFNE GKHQSRTIAS EARMLKK MF LKLRD

UniProtKB: Uncharacterized protein

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Macromolecule #4: Intraflagellar transport protein 46

MacromoleculeName: Intraflagellar transport protein 46 / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 37.97641 KDa
SequenceString: MDDSMDYPDR DGDDLDQFQG TARSQVVQNQ PHDEEVNLSE SESFAGADEP PAAPRDASLI ESHDMDEGPA APARTLSPTG YEAGKHAPG GIANSDEAPP GAYNAQEYKH LNVGEDVREL FSYIGRYKPQ TVELDTRIKP FIPDYIPAVG GIDEFIKVPR P DTKPDYLG ...String:
MDDSMDYPDR DGDDLDQFQG TARSQVVQNQ PHDEEVNLSE SESFAGADEP PAAPRDASLI ESHDMDEGPA APARTLSPTG YEAGKHAPG GIANSDEAPP GAYNAQEYKH LNVGEDVREL FSYIGRYKPQ TVELDTRIKP FIPDYIPAVG GIDEFIKVPR P DTKPDYLG LKVLDEPAAK QSDPTVLTLQ LRQLSKEAPG AKADMVGRLE HTDENKAKKI QQWIASINDI HKAKPAATVN YS KRMPEIE ALMQEWPPEV ETFLKTMHMP SGDVELDIKT YARLVCTLLD IPVYDDPVES LHVLFTLYLE FKNNPIFRQH MEM ENKLDG MSGGGGGMMG GGADVLGL

UniProtKB: Intraflagellar transport protein 46

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Macromolecule #5: Intraflagellar transport protein 56

MacromoleculeName: Intraflagellar transport protein 56 / type: protein_or_peptide / ID: 5 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 63.485508 KDa
SequenceString: MFYSKSRPQH AARTNVAQAA ANEKPKVPEL EEFLTKRDYL GAIALLSFRR HANRNDLKNL EWLAYCYFHY GEHDKALVIY KELLQHEDP DPMFFVYSAA CLYYMGMYKE AEEQALQGPK CALQTRILFH SAQRQGNDDK LMAYHGQLTD SIEDQLTLAS I HYQRSHFQ ...String:
MFYSKSRPQH AARTNVAQAA ANEKPKVPEL EEFLTKRDYL GAIALLSFRR HANRNDLKNL EWLAYCYFHY GEHDKALVIY KELLQHEDP DPMFFVYSAA CLYYMGMYKE AEEQALQGPK CALQTRILFH SAQRQGNDDK LMAYHGQLTD SIEDQLTLAS I HYQRSHFQ EATDIYKRLL LEHRDYLRST VVYVALCYCK LDYYDVSLEI LGVYLSAFPD SAIAVNLKAC NHFRMYNGKA AE AELKTLA ELSGGQHLDH DLIRHNLVVF RGGENALQVL PPLSDIPPEA RLNLVIHHLR HHEVGEAFGL IKDMEPSTPP EFI LKAVVH AMLGQVKGDP EHLKKAQQYY QLVGASASEC DTIPGRQCMA SCFFLLKQFE DVLVFLSSIK TYFLNDDDFN WNLG IAKAA TGKYKEAEET LLQIANDKYR NEYTYTSWLA RCYIMNGKAR LAWERYLQLE TNDESYQLLL LIANDCYKMG AFYFA CKAF DVLERLDPAP EYLEGKKGAA CGAFQMIVAG KEPKDLLRDV ISLLRGNNSA DQCEPIVAVM RKWAKNNGVK L

UniProtKB: Intraflagellar transport protein 56

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Macromolecule #6: Intraflagellar transport protein 81

MacromoleculeName: Intraflagellar transport protein 81 / type: protein_or_peptide / ID: 6 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 77.186484 KDa
SequenceString: MGDVSYIVDS LGLPPFSYQM SLLSFTEKGP QELLQLLSDV FSTISPKHQK VDVAKEVPDQ TADRLIGFLK IIKYRPNVQD PLLFRQLVA AGDRETLYQI LRWVVPQAQL LEKRAFVGYY LSFPDMPEEF NFDPDIMELK EEIKAMQQEF IELHKSSDAI K TLSKDTQA ...String:
MGDVSYIVDS LGLPPFSYQM SLLSFTEKGP QELLQLLSDV FSTISPKHQK VDVAKEVPDQ TADRLIGFLK IIKYRPNVQD PLLFRQLVA AGDRETLYQI LRWVVPQAQL LEKRAFVGYY LSFPDMPEEF NFDPDIMELK EEIKAMQQEF IELHKSSDAI K TLSKDTQA LKNKIKSLEE EKERLGEKVE RAKGAVDKLP DRSSYMEVCT NLRKQQDEEV NLSTAIQTQR ALQEKAEASY HR AAARLRE LQTSYQEGSA GKLLETLNED VKNLRAQVNE RYPKEVEKRQ KRQAALSEAL ASGISTEMDL QRLQHQATAL HNQ ITEIQE RKVAQDKARQ GDKAYLQLRQ AQQMATVSAR KKEELGAKQE RLQEKKTALT AQLDKLNAEG GGSGAVFSEE EWRT KYESM KSKLPIYKKM KKELGDLEAE VFVLAHTEEL LASQEGGLLE KVKRLEKQQG ISGFTETAQH LEKVSEAKSQ MDEEK GMTL IEISRTVEEI NNAINQRKQQ LAPQIKKLRS VRQDFAEFEA KYLEKKTAYD NVVATFEART SALEGEVSGL KAEVSE NET KYHMLHCQLH ITDQNIKKVT SGPAAERLRD KYEAKVKEAE DSTKALRDRQ REIKDTHSTG LSQIDIMNDM LRLLQLK LN LARGIAVDMS QYGGGGGAAG GANGGMAGQT YDTGSANVLQ L

UniProtKB: Intraflagellar transport protein 81

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Macromolecule #7: Intraflagellar transport protein 74

MacromoleculeName: Intraflagellar transport protein 74 / type: protein_or_peptide / ID: 7 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 71.648398 KDa
SequenceString: MDRPSSRGAL ALGAGGLGKA PTGGAVQQPD RPMTGQRGAA PAGPMRAPAG ASIIGAGPPG TAMRGGPGPA GGPPGTAYKR MGTASQRPG TGQQAAAAAA AARAGQQLQV ENRPITNHGV SGMKTAAAGV GRQVLDKNYF MNELRQKRME IAQVTSNMKS D LEALERKQ ...String:
MDRPSSRGAL ALGAGGLGKA PTGGAVQQPD RPMTGQRGAA PAGPMRAPAG ASIIGAGPPG TAMRGGPGPA GGPPGTAYKR MGTASQRPG TGQQAAAAAA AARAGQQLQV ENRPITNHGV SGMKTAAAGV GRQVLDKNYF MNELRQKRME IAQVTSNMKS D LEALERKQ AQYNSMDKRA SDLSKEVKIL QEALADYNTV LDKVGSQAPV YVIQQEFAAL KDRNEQQRKR VDEVLTERLN LE SKAKQAE SKMSEIQASM DQRLNSMPPS QRNEYTTLVA EQQQLQADSK RFEEVLDELD KALQASEGEL ARNPFKQRSL QLQ EQIRAL TEKKYELTEE ERQSKRSPEE LRADLMAKIK RDNTEVEQMT QQIRELQDQI KKMEERVKSL GGATSGAAAA EEKA NREKF EELLAKERDL NNFMDGFPSR KAAKMQEKQQ KEDGIVGVLE KMVKMQGIIG SNLPSQKKYK EMQDELEYKK MQLEN TQTT QERLKEELTM RRTELEKIDT LEDKIKLELT QLAERQEAME KEMGEFGSVE DIQRKANAAR ERMEGLRSVL LKRKDL LRS IVAERGLKFQ AKRAQLQDHN LQVQLEKMEA KLKNLSAGVF EMDEFIKAKE SETNYRQLAS NIAALVDDLN VHVKKAV V

UniProtKB: Intraflagellar transport protein 74

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Macromolecule #8: Intraflagellar transport protein 80

MacromoleculeName: Intraflagellar transport protein 80 / type: protein_or_peptide / ID: 8 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 85.782609 KDa
SequenceString: MRLKVKQSSA NVHSELTAAV GWNVWNELFT CSDDQTIHKW NMLGEPEQKV STLDAYFTDM HWYPVSSKKT QAGGTDVFAV ACTDGSVKI LSRTGRVEKS IEGHKGACIS LRWSYDGTAL ATAGEDGSVK IWSRNGMLRS TLAQADSPVY SIVWAYDCDQ L CYCTGSNV ...String:
MRLKVKQSSA NVHSELTAAV GWNVWNELFT CSDDQTIHKW NMLGEPEQKV STLDAYFTDM HWYPVSSKKT QAGGTDVFAV ACTDGSVKI LSRTGRVEKS IEGHKGACIS LRWSYDGTAL ATAGEDGSVK IWSRNGMLRS TLAQADSPVY SIVWAYDCDQ L CYCTGSNV VIKSLSSNAK QNAWKAHDGV VLKVDWSPIN HLIITGGEDC KYKVWDSFGR LLFQSGLFDY PVTSVAWAPS GE LFAVGGF NTLQLCDRMG WAYSKIHLND TGSIMTLSWT ADSTQLAGGG GSGGVVFGQV VDLALEDGKM QVTVVDDMRI VVN DILNEN ADELPEFRDR VIKVSLGYGY LIVATATQCH VYNTTNLGTP HIFDLKDTVT LLLQAERHFL LLDNSAGIQI YTYE GRQIC NPRFQGLRTE LLNAQMITLS NDTIAVLDQQ ASGTTVRFFD TAQGRPVGEP WQHTLEVKEI ALSQAGTIND RQLIV IDRN RDLYLLPVMK RHVAKLAAMC DSARWHDSTA MLSAMVDQRL CVWYYPSEVY VDKDLLAKTR YTKSDSDFGK SAQIQL FAG NRCLVRRSDG VLVSAATSPY PAVLYDMIRK QQWDKATRLC RFIKDPTMWA TLAAMAMAAK ELNTAEVAFA AIDEVDK TH FVRKVKQIPT EEGRNAELAV YRRKPEEGES ILLQAGLVFR AIKLNIKLFN WERALDLATQ HKQHQDTVLW YRQQFLKN A KLAESITRFM QMNESVVVDQ AAVKKKIEEE RIKESQRPGA KRYV

UniProtKB: Intraflagellar transport protein 80

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Macromolecule #9: Clusterin-associated protein 1

MacromoleculeName: Clusterin-associated protein 1 / type: protein_or_peptide / ID: 9 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 49.733469 KDa
SequenceString: MSFRELRSFT EVMKALGYPR LISMENFRVP NFELVADCLY WLVHRYYPGV EIADDISTES DRVKFLQSVA QVMLTKARMK LNIKRLYAA DGNAVKELLK LASLLYKATS KAGDMDDDAA EAIDVANSLK GFNPKEIKGL AAEIIKAGAG LYDALGQEPE L REHRARAV ...String:
MSFRELRSFT EVMKALGYPR LISMENFRVP NFELVADCLY WLVHRYYPGV EIADDISTES DRVKFLQSVA QVMLTKARMK LNIKRLYAA DGNAVKELLK LASLLYKATS KAGDMDDDAA EAIDVANSLK GFNPKEIKGL AAEIIKAGAG LYDALGQEPE L REHRARAV AGHVDTDFVE RSIREAIAQV EDNIRSLENQ MEDLERNEKT LDNKIEKKKQ ELERQEKRLS TLQSVRPAYM DE YERLQGE LNGLYQSYLD RFRNLEFLEN ELEAYYQAEQ EKMEAQERRL KKMQKRLKEE ELRILRGEQE VDEANMNFDD DDM DLSEDG EHEEAEEGED EDYGAGGQKA TMRRDQFGGQ GGGGGGGRQQ GGGRNVMGNL DGDFDDEASI DEDDNTDGGE VSRQ GSQAG GMGGGGGYGG GGGYMNGGGM DDDENNLIQD DDEDDNDF

UniProtKB: Clusterin-associated protein 1

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Macromolecule #10: Intraflagellar transport protein 57

MacromoleculeName: Intraflagellar transport protein 57 / type: protein_or_peptide / ID: 10 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 51.298414 KDa
SequenceString: MSSKRGGRSS LAKAPEEAVN GEAFAPEASP PPPGDDGDAG GEDGGAPAPP PPPATKGGPV AVGRSLEIQT TPDVCMEMLA DKLKLLNYE ADFCRKKKPY RKPLSRLYFA VPLANSSEQF FYFTSLATWL LGLAGVELPA PKEFDDPNLT CQNILGAVKK L GFAPPSYH ...String:
MSSKRGGRSS LAKAPEEAVN GEAFAPEASP PPPGDDGDAG GEDGGAPAPP PPPATKGGPV AVGRSLEIQT TPDVCMEMLA DKLKLLNYE ADFCRKKKPY RKPLSRLYFA VPLANSSEQF FYFTSLATWL LGLAGVELPA PKEFDDPNLT CQNILGAVKK L GFAPPSYH PTKLTVGNGK EVVGVLDGLV DFVLERRHHK YSRPAYGNDG QPEEGVQLDD EAEAAAMEGA DELAMPAQNQ AD DDEEEEG VYVDPGRGDA AGPGTGASAA MDAEKAVLVS KVDPTLWKIE LERVAPKLRI TIAADSKDWR SHLDEAHQHK EVI SKAWPD SKTSLERLRA DLNGTLEKLQ TREKFLNEQA ESLMQQYRAA RTTFTDVQET YNRKTEAVAD RNQEMHRIGE TLEE VKAMM DEKGSNIADA TPVARIKTAI KQLNKELHDM EVRIGVVSHT LLQLSLRNKR LLQAQAALSD EEED

UniProtKB: Intraflagellar transport protein 57

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Macromolecule #11: Intraflagellar transport particle protein IFT20

MacromoleculeName: Intraflagellar transport particle protein IFT20 / type: protein_or_peptide / ID: 11 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 15.658664 KDa
SequenceString:
MDAVDRGVYF DEDFHVRILD VDKYNASKSL QDNTNVFINN IQNMQGLVDK YVSAIDQQVE RLEAEKLKAI GLRNRVAALS EERKRKQKE QERMLAEKQE ELERLQMEEQ SLIKVKGEQE LMIQKLSDSS SGAAYV

UniProtKB: Intraflagellar transport particle protein IFT20

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Macromolecule #12: IFT54

MacromoleculeName: IFT54 / type: protein_or_peptide / ID: 12 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 54.437715 KDa
SequenceString: MCDNWQATID TLQGASPVFD KPKLSQKLLE KPPFRFLHDV VTAVQQATGF APGLYQGDEL DGKAIQEKDA KVAYLKKIIE VVSMVLGEQ CPARPNKIVA GLEPENTNIF LQMLGRACQK GNGAKAVQKV LGGGGAEPAP AKEEAPPPEK KPEKKEKKEE K PAEKSRAE ...String:
MCDNWQATID TLQGASPVFD KPKLSQKLLE KPPFRFLHDV VTAVQQATGF APGLYQGDEL DGKAIQEKDA KVAYLKKIIE VVSMVLGEQ CPARPNKIVA GLEPENTNIF LQMLGRACQK GNGAKAVQKV LGGGGAEPAP AKEEAPPPEK KPEKKEKKEE K PAEKSRAE ASPARKKAAE PEAEKKSSSK SSSRTKEEPP AKAPAKKKEE PAPEKPSKSK AAPAAEEAPP PPPPPAAEPP AR SASPGGE DPLNKSGSAA PKFQRPTSAR KAPPRVPQPQ QPTMLAGTGI RPGTATRRPN EPKPTDSKVT KPVAVFTDNA KDN SDDEVE VVHEQTPVLS GGANMTGEQG VLVKDILAAE KGLKKAGVDA TADNADTSDQ GSTGIILKRL GGKAAGAGAA AAGP RAHDP SSVRELVQKL CHSSTPLAKS MDYLQEDIEN MRKEYKFWLT EKRMYQDELA RELRLQGEAA NVDAQLADLD GQIKQ ARDR IIGMKGQILR NDETLGKLLA MATAGR

UniProtKB: Uncharacterized protein

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Macromolecule #13: Intraflagellar transport protein 172

MacromoleculeName: Intraflagellar transport protein 172 / type: protein_or_peptide / ID: 13 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 197.851172 KDa
SequenceString: MQLRYFKSIL PPADQYQKIT SLTWAPNNSR LAAVSTDKVV YLFDENGEKR DKFKTKAAEA NNPNTYIIRA MAFSPDSTKL AIAQSDNIV FIYRLVDPDT GAEKKSICNK FPQACAVTSL VWPKDRPNEV VFGLADGKVR LGMLKNNKSY TCYAHPENSY V VALASSLN ...String:
MQLRYFKSIL PPADQYQKIT SLTWAPNNSR LAAVSTDKVV YLFDENGEKR DKFKTKAAEA NNPNTYIIRA MAFSPDSTKL AIAQSDNIV FIYRLVDPDT GAEKKSICNK FPQACAVTSL VWPKDRPNEV VFGLADGKVR LGMLKNNKSY TCYAHPENSY V VALASSLN GQNVISGHMD GAIWKFNFPA EEGGTPTSSQ LVVHSCVPYS LGWGSCIAAA GNDNRVVFYD LNGREIRSFD YS NNDEVRE FTTCAFNPSG DTVVFGTYNR FYMYTFNIQR NDWEEAGHKQ IDNFYAVSAA SWKPDGSKMT VGSMTGAVDM YDA CVKRHM YKGKFEFTYV SKSAVIVKTL KTGMRIVLKS VYGYEIEKIN IYHDRYLIAR TTYTLLMGDL DTCKLSEIPW DSDG SEKFH FENERVCMVH YAGELHIVEY GRNDVLGTCR TEHMNPYLIS AVVQEARGIA SESKKLAYLI DLQTVRIQDL MAPVG STLA TVNHDTKVDW LELNQRGTHL LFRDKKRHLH LFSLSGQERT TLLNYCQYVQ WVPGSDVIVA QSRNNLCVWY SVNKPD NVT MFPIKGEVVD IERHNHRTEV IVDEGINTVS YALDEALIYF GAALEDQDYE RAVQTLEPLE LTPETEAQWM QLAEQAL AT NQLVIAERCY AALGDIAKSR FLHKVVKKAQ QAAKEFGGDG TDAWSVRAMM AQLNKQWPVS ESLLLAQGKV DDAITLYQ D NHRWEDAIRV ADSTHHANAA ALKQQYLTWL LETGQEEQAG AVKEREGDYL AAIGLYLKGG LPGRAAQVVM SVHNVNWDP ALLDSILASL AKAGLYERAG ELYEHMSRSS EAMQSYRRGH AYRKAIDLAR REFPAEVIII EEEWGDWLVT QKQMDAAINH FIESGATLK AIKAAIDCRQ FAKAAGIIEV LDPREAMPYF RRIAQHYETT GALEEAERYY IRADMARDAV EMYSRAGKWE A AQRVARGY LTESEMRAFY RAKAAEFEAA HKLKEAEKAY LAAGGDDVDK AIAMYKRNKM YDQMIRLVTQ YRKEKVPEAH TL IAQQLEV EGNLREAEKH FVEAKDWKSA VQMYRQVNQW EDALRVAKVY GGVNASKQVA YAWALTLGGD DGAQLLKKMG LLD HAIEYA VESGAFAQAF EMTRAGAKHK LPEVHLKYAM FLEDEGRFAE AEAEFISAGK PKEACDMYMH NQDWDAAMRI AERY DPTMV SEILVSQARV AVERKQWLPA EGLFIKAKRP EAALKMYRDA RMWNDALRVA EQYLPTKVAE VQMELLSGQG AGGGS GGAS ADAVINKARG FERNNDYARA IETYLSLTAQ DTSNQDQLEH CWGQAAQLAI NYQRHRMKDV VNTVSERLQE IGRHQA AGE LHESIDDAQG AIRAYCAGRL WDKARTLAGT NPTFSRYIED QYNNYLLQNQ QADELASRGG QHAQQAIEMY VARDEWA KV HELAAQQGPE VASNYALKHA ERRFKQGDYA QAAQVFAQHG ITAQPQYFEL YKSIAQGVLH ASQGDRNPVA EKSLRDMM Y RLVNVLRSGG GAGKYKVDTD AFQNYYLAAH YLTCAAAAKE QGLKDIAAMN LTSVLRYVGP TIPADRAFYE AGLAWYEAG RKNMAFVMLN RFLDLSDAMD EPDSSAAVIE NADFSDTDIP YDFTIPERAY CTESQREDVR NLVLEISMDR SSDQSLALKA CEHCGKPTY EANLTCHFCK KKYDPCVVTG YPIQSYDRVV FKNNGPELNA IRDMWNKWVE AFGTDPVTGM QAAPMY

UniProtKB: Intraflagellar transport protein 172

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statehelical array

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Sample preparation

BufferpH: 7 / Details: TAP (Tris-Acetate-Phosphate) Media
GridModel: Quantifoil R3.5/1 / Material: GOLD
VitrificationCryogen name: ETHANE
DetailsChlamydomonas reinhardtii cells applied to quantifoil grids and plunge frozen; cilia project out from cell bodies and traverse holes in the carbon film.

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 2.6 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 5.0 µm / Nominal defocus min: 2.5 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 9.9 Å / Resolution method: FSC 0.143 CUT-OFF / Software: (Name: RELION (ver. 3.1.3), Warp) / Number subtomograms used: 18216
ExtractionNumber tomograms: 600 / Number images used: 18216
Final angle assignmentType: MAXIMUM LIKELIHOOD

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Atomic model buiding 1

RefinementProtocol: FLEXIBLE FIT
Output model

PDB-8bd7:
IFTB1 subcomplex of anterograde Intraflagellar transport trains (Chlamydomonas reinhardtii)

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