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- EMDB-22475: Composite cryo-EM density map of the radial spoke 1 isolated from... -

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Basic information

Entry
Database: EMDB / ID: EMD-22475
TitleComposite cryo-EM density map of the radial spoke 1 isolated from Chlamydomonas reinhardtii
Map datacomposite map
Sample
  • Complex: radial spoke 1
    • Protein or peptide: x 19 types
  • Ligand: x 1 types
Function / homology
Function and homology information


radial spoke head / radial spoke / positive regulation of cilium-dependent cell motility / kinocilium / cilium movement involved in cell motility / cilium movement / motile cilium assembly / axoneme assembly / dynein complex / Set1C/COMPASS complex ...radial spoke head / radial spoke / positive regulation of cilium-dependent cell motility / kinocilium / cilium movement involved in cell motility / cilium movement / motile cilium assembly / axoneme assembly / dynein complex / Set1C/COMPASS complex / UTP biosynthetic process / ciliary plasm / CTP biosynthetic process / Oxidoreductases / motile cilium / GTP biosynthetic process / nucleoside diphosphate kinase activity / axoneme / chaperone cofactor-dependent protein refolding / cilium assembly / microtubule-based process / protein peptidyl-prolyl isomerization / centriole / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / cilium / unfolded protein binding / protein folding / mitotic cell cycle / protein-folding chaperone binding / microtubule / oxidoreductase activity / cytoskeleton / calmodulin binding / phosphorylation / calcium ion binding / ATP binding / membrane / metal ion binding / cytosol / cytoplasm
Similarity search - Function
Nucleoside diphosphate kinase 6 / Radial spoke protein 14 / IQ and ubiquitin-like domain-containing protein / Radial spokehead-like protein / Radial spoke 3 / Radial spokehead-like protein / Radial spoke protein 3 / Sdc1/DPY30 / Dpy-30 motif / Dpy-30 motif ...Nucleoside diphosphate kinase 6 / Radial spoke protein 14 / IQ and ubiquitin-like domain-containing protein / Radial spokehead-like protein / Radial spoke 3 / Radial spokehead-like protein / Radial spoke protein 3 / Sdc1/DPY30 / Dpy-30 motif / Dpy-30 motif / Possible plasma membrane-binding motif in junctophilins, PIP-5-kinases and protein kinases. / Dynein light chain, type 1/2, conserved site / Dynein light chain type 1 signature. / cAMP-dependent protein kinase regulatory subunit, dimerization-anchoring domain / Regulatory subunit of type II PKA R-subunit / RIIalpha, Regulatory subunit portion of type II PKA R-subunit / MORN motif / HSP40/DnaJ peptide-binding / MORN repeat / Chaperone DnaJ, C-terminal / DnaJ C terminal domain / Dynein light chain type 1 / Dynein light chain, type 1/2 / Dynein light chain superfamily / Dynein light chain type 1 / Cytochrome b5 family, heme-binding domain profile. / Cytochrome b5-like heme/steroid binding domain / Cytochrome b5-like heme/steroid binding domain superfamily / Cytochrome b5-like Heme/Steroid binding domain / Cytochrome b5-like Heme/Steroid binding domain / EF hand / Nucleoside diphosphate kinase, active site / Nucleoside diphosphate kinase (NDPK) active site signature. / DnaJ domain / Nucleoside diphosphate kinase / Nucleoside diphosphate kinase-like domain / Nucleoside diphosphate kinase / NDK / Nucleoside diphosphate kinase-like domain superfamily / IQ calmodulin-binding motif / DnaJ molecular chaperone homology domain / dnaJ domain profile. / Chaperone J-domain superfamily / DnaJ domain / Cyclophilin-type peptidyl-prolyl cis-trans isomerase / Armadillo/plakoglobin ARM repeat profile. / Short calmodulin-binding motif containing conserved Ile and Gln residues. / Armadillo/beta-catenin-like repeats / Armadillo / IQ motif profile. / NADP-dependent oxidoreductase domain / Aldo/keto reductase family / NADP-dependent oxidoreductase domain superfamily / Cyclophilin-type peptidyl-prolyl cis-trans isomerase domain profile. / Cyclophilin-type peptidyl-prolyl cis-trans isomerase domain / Cyclophilin type peptidyl-prolyl cis-trans isomerase/CLD / Cyclophilin-like domain superfamily / IQ motif, EF-hand binding site / EF-hand domain pair / EF-hand, calcium binding motif / EF-Hand 1, calcium-binding site / EF-hand calcium-binding domain. / EF-hand calcium-binding domain profile. / EF-hand domain / Armadillo-like helical / EF-hand domain pair / Armadillo-type fold
Similarity search - Domain/homology
IQ and ubiquitin-like domain-containing protein / Cytochrome b5 heme-binding domain-containing protein / EF-hand domain-containing protein / Radial spoke protein 14 / Peptidyl-prolyl cis-trans isomerase / Radial spoke protein 4 / Radial spoke protein 16 / Uncharacterized protein / Predicted protein / Flagellar radial spoke protein 3 ...IQ and ubiquitin-like domain-containing protein / Cytochrome b5 heme-binding domain-containing protein / EF-hand domain-containing protein / Radial spoke protein 14 / Peptidyl-prolyl cis-trans isomerase / Radial spoke protein 4 / Radial spoke protein 16 / Uncharacterized protein / Predicted protein / Flagellar radial spoke protein 3 / Flagellar radial spoke protein 4 / Flagellar radial spoke protein 6 / Flagellar radial spoke protein 1 / Radial spoke protein 11 / Radial spoke protein 10 / Radial spoke head protein 9 homolog / Flagellar radial spoke protein 5 / Dynein 8 kDa light chain, flagellar outer arm / Nucleoside diphosphate kinase 6 / Flagellar radial spoke protein 2
Similarity search - Component
Biological speciesChlamydomonas reinhardtii (plant)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsGui M / Ma M / Sze-Tu E / Wang X / Koh F / Zhong E / Berger B / Davis J / Dutcher S / Zhang R / Brown A
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS) United States
CitationJournal: Nat Struct Mol Biol / Year: 2021
Title: Structures of radial spokes and associated complexes important for ciliary motility.
Authors: Miao Gui / Meisheng Ma / Erica Sze-Tu / Xiangli Wang / Fujiet Koh / Ellen D Zhong / Bonnie Berger / Joseph H Davis / Susan K Dutcher / Rui Zhang / Alan Brown /
Abstract: In motile cilia, a mechanoregulatory network is responsible for converting the action of thousands of dynein motors bound to doublet microtubules into a single propulsive waveform. Here, we use two ...In motile cilia, a mechanoregulatory network is responsible for converting the action of thousands of dynein motors bound to doublet microtubules into a single propulsive waveform. Here, we use two complementary cryo-EM strategies to determine structures of the major mechanoregulators that bind ciliary doublet microtubules in Chlamydomonas reinhardtii. We determine structures of isolated radial spoke RS1 and the microtubule-bound RS1, RS2 and the nexin-dynein regulatory complex (N-DRC). From these structures, we identify and build atomic models for 30 proteins, including 23 radial-spoke subunits. We reveal how mechanoregulatory complexes dock to doublet microtubules with regular 96-nm periodicity and communicate with one another. Additionally, we observe a direct and dynamically coupled association between RS2 and the dynein motor inner dynein arm subform c (IDAc), providing a molecular basis for the control of motor activity by mechanical signals. These structures advance our understanding of the role of mechanoregulation in defining the ciliary waveform.
History
DepositionAug 17, 2020-
Header (metadata) releaseDec 16, 2020-
Map releaseDec 16, 2020-
UpdateJan 27, 2021-
Current statusJan 27, 2021Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.006
  • Imaged by UCSF Chimera
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  • Surface view colored by cylindrical radius
  • Surface level: 0.006
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-7jtk
  • Surface level: 0.006
  • Imaged by UCSF Chimera
  • Download
  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-7jtk
  • Imaged by Jmol
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_22475.png

Downloads & links

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Map

FileDownload / File: emd_22475.map.gz / Format: CCP4 / Size: 669.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationcomposite map
Voxel sizeX=Y=Z: 1.09 Å
Density
Contour LevelBy AUTHOR: 0.006 / Movie #1: 0.006
Minimum - Maximum0.0 - 0.07739167
Average (Standard dev.)8.5046835e-05 (±0.0009487174)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions560560560
Spacing560560560
CellA=B=C: 610.4 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.091.091.09
M x/y/z560560560
origin x/y/z0.0000.0000.000
length x/y/z610.400610.400610.400
α/β/γ90.00090.00090.000
start NX/NY/NZ1081340
NX/NY/NZ13584349
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS560560560
D min/max/mean0.0000.0770.000

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Supplemental data

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Mask #1

Fileemd_22475_msk_1.map
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Mask #2

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Mask #3

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Mask #4

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Mask #5

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Mask #6

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Mask #7

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Mask #8

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Additional map: #6

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Additional map: #5

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Additional map: #4

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Additional map: #3

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Additional map: #2

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Additional map: #8

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Additional map: #7

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Additional map: #1

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Sample components

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Entire : radial spoke 1

EntireName: radial spoke 1
Components
  • Complex: radial spoke 1
    • Protein or peptide: Flagellar radial spoke protein 1
    • Protein or peptide: Flagellar radial spoke protein 2
    • Protein or peptide: Flagellar radial spoke protein 3
    • Protein or peptide: Flagellar radial spoke protein 4
    • Protein or peptide: Flagellar radial spoke protein 5
    • Protein or peptide: Flagellar radial spoke protein 6
    • Protein or peptide: Flagellar radial spoke protein 7
    • Protein or peptide: Flagellar radial spoke protein 9
    • Protein or peptide: Flagellar radial spoke protein 10
    • Protein or peptide: Flagellar radial spoke protein 11
    • Protein or peptide: Flagellar radial spoke protein 12
    • Protein or peptide: Flagellar radial spoke protein 14
    • Protein or peptide: Flagellar radial spoke protein 16
    • Protein or peptide: Dynein 8 kDa light chain, flagellar outer arm
    • Protein or peptide: Unknown protein
    • Protein or peptide: Flagellar radial spoke protein 23
    • Protein or peptide: Cytochrome b5 heme-binding domain-containing protein
    • Protein or peptide: Uncharacterized protein
    • Protein or peptide: Predicted proteinProtein structure prediction
  • Ligand: PHOSPHATE IONPhosphate

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Supramolecule #1: radial spoke 1

SupramoleculeName: radial spoke 1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#19
Source (natural)Organism: Chlamydomonas reinhardtii (plant) / Strain: CC-1690 / Organelle: cilia

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Macromolecule #1: Flagellar radial spoke protein 1

MacromoleculeName: Flagellar radial spoke protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 87.873445 KDa
SequenceString: MALGPFVLPF RGDQYSFGIN FKSSPEEKLN FDLSCVAFDV KGQLHDTLHA RKPTALDGAL VKGFEKQALP EETVQVEGDD VIYMFPKKF ERQVEVLLFV ASAPSIPGKK HDLDSSSKLE FAVSYSDVGG QAFNQSFDLK PLAAQGGVSS IIVAVMYLQA E GGWTLRSV ...String:
MALGPFVLPF RGDQYSFGIN FKSSPEEKLN FDLSCVAFDV KGQLHDTLHA RKPTALDGAL VKGFEKQALP EETVQVEGDD VIYMFPKKF ERQVEVLLFV ASAPSIPGKK HDLDSSSKLE FAVSYSDVGG QAFNQSFDLK PLAAQGGVSS IIVAVMYLQA E GGWTLRSV GDCHPFDSPG LIVPELKQTI LNLRDHHGVQ LDAADAIQAI DPAERVPVTR QFQDQSLDEA SAGRAAEPAP VK KLRIDLS WTFWPPPPPT EEGEEPPEEP ALEYNLVMYN KDGEEVQSIS TGNREATGAR AGRPEPEEDE EEEKEEEKEE PEE GEEGEE GEGGEPKEPP PPPPAPKVDP YEFKERDVIY LDVPDLPAEV RSMVLLVTNY DEENGFTRVR TVRCRLVDVS NGEA PLPGS KAAVAAAAAA AEQGLAAPPN PERVLADYGV LSKYEDDKAT TQVALMKLYK EYADSAFNVF RGAGVDNVAA FIGQE PDTI INQLKAYLEA TKKQKAAEAA AAAAAEESGE EITADPKPHV WRFRALGLNF GGDSLEAIEH DLKNLFAFDG DLAPGA ARD SDTSRSSFPN GDTYFGSYAD DVKHGPGLYA FATGAGYAGE YAGGKRHGRG VMVFPDGGTY VGEFVADKFE GQGQYRY PD GSVYTGSWAA GQKHGPGVYW DTARGCLRGE WKKGLLVGKG TYEQPALRFE GEFVRGMPAG TATYTLTGHR TLDMPCFA A QHIQAEEGPT LALPCAYGIP PGSGDEPQLD EEGQPIEDTD KPPLPAHPKY EGLTFTAEQL PGAAPDTVFP PEEGKPVPI TAVPAFSVST GLVA

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Macromolecule #2: Flagellar radial spoke protein 2

MacromoleculeName: Flagellar radial spoke protein 2 / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 77.445297 KDa
SequenceString: MAPTQAGHDT AYLKETVGEA LARGCAAAIS AQPNDPVEYL GLWLLKYVKN AEVEGNFYRE RQQDLQKKKD RLVKEAQSEQ AAKSVALTR KEAADALALV TAEPRELLEA AVKLVKQHTA AGAAYAAVVA EPEEPDWVAP EDDEAAAVET EDEAAGGAAL A EGEEPPPE ...String:
MAPTQAGHDT AYLKETVGEA LARGCAAAIS AQPNDPVEYL GLWLLKYVKN AEVEGNFYRE RQQDLQKKKD RLVKEAQSEQ AAKSVALTR KEAADALALV TAEPRELLEA AVKLVKQHTA AGAAYAAVVA EPEEPDWVAP EDDEAAAVET EDEAAGGAAL A EGEEPPPE PEPEPEAAPE DGEGDAPAPK IPRPVDYSKK YFAYVAASAG QEHVLEADLY RPAPPPEDAD EDFKPEPLPY SF RVLDEKL PMLYVPNVAA EERVKFFRKF PKIGSYQACG VALPASGEFK ALLAADTLFP EGSGQPLSAD DRDFVWEVSQ SLS RALEAV QARAAEALAA TSAAEAVEEL KAKVAELREQ AAAEAAAAAP PPPAEGEEGE GEAPPAEEEP PAEEEAEEEE EEAE EGAEE GAEEGEEGEE APPKPKKKKK VFNPIPGLQA AIEKLTAAAE AATEADARAQ AAVALEKQAL DEVVALASSH SDATL SSLR NMLSVPQGTY HVVKALLHLL GRPAASFSTW KRAHSHFSPR LFEDMAAYDA TAERDMAVWG RVRSCYKAAP AAKKLD AEM PNTLFGSVAL MYIKQVRRVA RKAVLQRELA AKLAKAQQDL ADKQAALVEA ERVKAEREAE EARLAAEAEA AAAAEAE AA ARAAAEAEAA AAAEAAAEAA AEAAAAAAEA AAEAGEGEAV AEREAAPAEA EAAPAEGEAA PPAEGEGEAQ PAQEGSNS S SSSSDSSSSE ESKAAAE

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Macromolecule #3: Flagellar radial spoke protein 3

MacromoleculeName: Flagellar radial spoke protein 3 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 56.856688 KDa
SequenceString: MVQAKAQQQL YTHAAEPKAV QQRRAKYRED ETTQTLPTAN IMFDRRVVRG NTYAARILPA DATQTQTKGP SPASTKKRTT RTLPPRTPE AVDGRRHIDI QTDVYLEELT DTVPEADTST QTDAFLDRPP TPLFVPQKTG TDAITQIENG DLFDFDFEVE P ILEVLVGK ...String:
MVQAKAQQQL YTHAAEPKAV QQRRAKYRED ETTQTLPTAN IMFDRRVVRG NTYAARILPA DATQTQTKGP SPASTKKRTT RTLPPRTPE AVDGRRHIDI QTDVYLEELT DTVPEADTST QTDAFLDRPP TPLFVPQKTG TDAITQIENG DLFDFDFEVE P ILEVLVGK VLEQGLMEVL EEEELAAMRA HQEHFEQIRN AELVATQRME AAERRKLEEK ERRMQQERER VERERVVRQK VA ASAFARG YLSGIVNTVF DRLVSSGYIY DPVMREVETA FMPWLKEQAI GYLARGVVAR RVVDKLVEDA AAALAANRST LAD KAASTA ATVDAWAERQ AKMEAELQGK ELEAVRRRPT FVLRELKPAV ASADAVEAAA AELTAQAEEA ANAKWEADKA EAAE KARAE AEAAAEEQKA LLEELAATAA AEAEERGEEP PAEPPSLPDG VEPVDVEAEV AKAVEAVPKP PVKEVTDIDI LSYMM DKGA ITKDAIIQAL AVHALGDKAY TNHPAFAEAE GA

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Macromolecule #4: Flagellar radial spoke protein 4

MacromoleculeName: Flagellar radial spoke protein 4 / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 49.84377 KDa
SequenceString: MAAVDSVAQA LAYLQVHSPQ DGTSMYDHLV KLVSKVLEDQ PKNAVDLLET SLLVKKSTFD PKESSPLVPI PVAPDATQTQ AAVSIFGDP ELPINPATGE PVPADPPNEF EAENMLGAAA VLDCLGVGLG RELGVNIALA AKRIGEDPKL AVRSVRFFGK F LGLYSDYF ...String:
MAAVDSVAQA LAYLQVHSPQ DGTSMYDHLV KLVSKVLEDQ PKNAVDLLET SLLVKKSTFD PKESSPLVPI PVAPDATQTQ AAVSIFGDP ELPINPATGE PVPADPPNEF EAENMLGAAA VLDCLGVGLG RELGVNIALA AKRIGEDPKL AVRSVRFFGK F LGLYSDYF VFEVAFKKEA AKEAAPAAPA PERVEGEAAS SSAPEVPVEE PGKGANKFTY LVCSSLGGPL TRLPDVTPAQ VK ASRRIKK LLTGRLTSHV STYPAFPGNE ANYLRALIAR ISAATVVAPS DLFSLNDETG ELERAEDWEP PAGREMAAPT AWV HVRPHL KSQGRCEVHK RELPEDADED EFYNEDELEE GPDLLAALEE DAQLPGEQAA WTPIYSSASE AVKTQAGGLR SLVW PGAVC GGRGSEWTCV YVGWGVKNAP FVPLPPPPVA QEFAWGEVET QELELKPAPP PPEEEAEADE

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Macromolecule #5: Flagellar radial spoke protein 5

MacromoleculeName: Flagellar radial spoke protein 5 / type: protein_or_peptide / ID: 5 / Number of copies: 2 / Enantiomer: LEVO / EC number: Oxidoreductases
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 55.403551 KDa
SequenceString: MSEPGEEPVA APAGPAPDPV LNELYGSERP AVELLPGVPL SPIVNSCWLP ADAKAMLAES WIPVPPEDAG EEAGPPPPAF EAAAPEYNE LVRRLAKTAP FRKWNELTIQ AKQLEQEVAG LKGPDAEAKQ AELENVKVQI ADAEAAVAEV KQSFSDDPLS L TGWMQALT ...String:
MSEPGEEPVA APAGPAPDPV LNELYGSERP AVELLPGVPL SPIVNSCWLP ADAKAMLAES WIPVPPEDAG EEAGPPPPAF EAAAPEYNE LVRRLAKTAP FRKWNELTIQ AKQLEQEVAG LKGPDAEAKQ AELENVKVQI ADAEAAVAEV KQSFSDDPLS L TGWMQALT DLADGGMTTF EVSGQGWPYC SLRQLFGEMP SAAPPAGFFD GVERVLGTFK RRYEKERGPG SVQLMLKLAP NV FSDAWST GGAPAAVAAV EAYVERARAN VFGPDGGVTP EGVPEPLDLV QLVWWDFAAA DPLPVLKALQ RMATDQLQVD EDS GEVSVS EPKKIRGIGL VDFPADRLKA AIQAGVPITC VQVEHSVLVR SAQPVLDLCA KYGIKVLARG GTLGGLLSAK YLGA PPPDP VRGDADLDSV PGCLDAVNNV GGWARLQAAL AVIKGIADKH GVKPETVALR WQIDAGCFPL VTTRWSSRVW RQFGY EGWS SFEVSGGRPG VDGPLFQVES FLDVEDVRAL AGLAAVH

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Macromolecule #6: Flagellar radial spoke protein 6

MacromoleculeName: Flagellar radial spoke protein 6 / type: protein_or_peptide / ID: 6 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 48.884551 KDa
SequenceString: MAADVGQALA FLQQVKTTQG ASIYEGLKAA LAKVLEDRPV NAVEALETSV LSTPPAANLS VPLVPAASAA AAAAAVAKAS LFGDPEPVL DPESGEPIDP DAPNEFECED VEGDGDLLDG LGVGLGRQEM YAAMLAVKRL GEDAKRGVST VRFFGKFFGT Q ADYYVFET ...String:
MAADVGQALA FLQQVKTTQG ASIYEGLKAA LAKVLEDRPV NAVEALETSV LSTPPAANLS VPLVPAASAA AAAAAVAKAS LFGDPEPVL DPESGEPIDP DAPNEFECED VEGDGDLLDG LGVGLGRQEM YAAMLAVKRL GEDAKRGVST VRFFGKFFGT Q ADYYVFET TLQSNPDMPE APEGTIPLEP YGEGVNAYIY FVSNTLGGPL QQLPYVTPEQ IKASRLLRRY LTGRLDAPVS AF PAFPGNE ANYLRALIAR ISAATVCCPR GFFTADDDSA ELSANDEWVP LKGREMALPV NWSHRYAHLK GQGRTVTHKR DPP DEEEEP EKNFWTAEEM EAGPPPLATL DTDAPLPAAT GDKVPPPAWS PVFASASVTT RNQVAGVRSN RWPGAVCACA GRHF TSMYV GWGIKAGGEW SPCPPPPPVP QWGAPAAGVE GGQQLLLECN DLPPKPAPPE EEDE

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Macromolecule #7: Flagellar radial spoke protein 7

MacromoleculeName: Flagellar radial spoke protein 7 / type: protein_or_peptide / ID: 7 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 54.781898 KDa
SequenceString: MSSTYQKPIT IPGDFPAILK AFTREILRAQ PSNIYEFGAR YFSGLQGQAE PEGLGRAAPV EPASTSHAAT SKATDVEETA VMFDIAALT PAELEPILMK LFIEADADRS GFLDRHEFTA VLRNANLKLS DRQIRQILAE ADENDDDVIQ YKEFLPIMVD I LQSIKAKE ...String:
MSSTYQKPIT IPGDFPAILK AFTREILRAQ PSNIYEFGAR YFSGLQGQAE PEGLGRAAPV EPASTSHAAT SKATDVEETA VMFDIAALT PAELEPILMK LFIEADADRS GFLDRHEFTA VLRNANLKLS DRQIRQILAE ADENDDDVIQ YKEFLPIMVD I LQSIKAKE QAKAMMHGVE TMVRTEVETM LLHGLPQEEL QALMLKVFKK ADADGSGQLN RHEFKEALKA AELGLTRKDI NL ILSHIDV DRDGLVSYEE FIPVCFQVLV ERFKDEIVVN DILGNADELQ QMLLGAFRDA DPDNTGLLSQ RQVKSIFKEL SYK ALGLTT LQMVSLISQA PTTPDGMVQY IQFVPQAASI IRSMYDVETM KGRMHAIKAV AEAGGIAALG ALDLDQLRGV LEQA FQRVD TEGAGQLTLP QVTQVLDGLN SLAPDANLAL SDQHMKAMFA AIDADESGTV DWTELVNFIC DALEHIEREA YVANM RDGG AGGAGEAEAS PGDEEA

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Macromolecule #8: Flagellar radial spoke protein 9

MacromoleculeName: Flagellar radial spoke protein 9 / type: protein_or_peptide / ID: 8 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 29.572348 KDa
SequenceString: MVQLEPNITL VLKHLASCGA VVSAEQQAAL DHSIPIKRIE AGLRSLTLWG RLTTLNGKDY LVAEGYNVAS SKEGAAVYET KYFYSQDGA RWSDLQPVDS ETATRCARIK GMLSGDPAKN YELEEKDPNA PEPSPEAEEE VKPLVFQIPE LAVLRCRVDA I ATATSVIP ...String:
MVQLEPNITL VLKHLASCGA VVSAEQQAAL DHSIPIKRIE AGLRSLTLWG RLTTLNGKDY LVAEGYNVAS SKEGAAVYET KYFYSQDGA RWSDLQPVDS ETATRCARIK GMLSGDPAKN YELEEKDPNA PEPSPEAEEE VKPLVFQIPE LAVLRCRVDA I ATATSVIP TDSTILNAAS QVVPNRLFAG AAYPEKLESY QHRFSLPGSG VTLSQDLRGT WAVQYDAFKG VAQVRSLLFP GY FFYYAAN ELTWGSLYVG DGLRNNDLIF ML

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Macromolecule #9: Flagellar radial spoke protein 10

MacromoleculeName: Flagellar radial spoke protein 10 / type: protein_or_peptide / ID: 9 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 23.553943 KDa
SequenceString: MADDELPPQP VWEGPLDEDG KPHGLGKMEY PPPPMGEDDE EEKPGDKFEG TMEHGVRTGK GTYTWGVSGA VYTGDYVNGK KHGKGKMVY PDKGVYEGDW VEDVMQGQGT YTYPNGDIYQ GAFWAGKRHG KGMYHYKGPC CQLVGDWADG GFTYGRWVYA D GSMFMGKF ...String:
MADDELPPQP VWEGPLDEDG KPHGLGKMEY PPPPMGEDDE EEKPGDKFEG TMEHGVRTGK GTYTWGVSGA VYTGDYVNGK KHGKGKMVY PDKGVYEGDW VEDVMQGQGT YTYPNGDIYQ GAFWAGKRHG KGMYHYKGPC CQLVGDWADG GFTYGRWVYA D GSMFMGKF GGAAADSKPT AGSYFYSSSS LVQEGHFAKD GSWVGHRDPA VGKEFSVA

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Macromolecule #10: Flagellar radial spoke protein 11

MacromoleculeName: Flagellar radial spoke protein 11 / type: protein_or_peptide / ID: 10 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 21.504635 KDa
SequenceString: MDVEPIFCAE QIVIPHNLAD ILKAYTKEVI RRQPTDLIAF SAKYFTNLAN VASGVSNSSA PAKEQLRQVY TRGGSGGATL TESQVTGLC QQAGIADAVV AKVMEVGAFT PAAVDLSKFV FLCLAMSCED FNRVCMGVFD VFSDNGSLPA QDLLTLIAHL G PDMDPEVT ...String:
MDVEPIFCAE QIVIPHNLAD ILKAYTKEVI RRQPTDLIAF SAKYFTNLAN VASGVSNSSA PAKEQLRQVY TRGGSGGATL TESQVTGLC QQAGIADAVV AKVMEVGAFT PAAVDLSKFV FLCLAMSCED FNRVCMGVFD VFSDNGSLPA QDLLTLIAHL G PDMDPEVT PAFLDAVAAE LPAGGGAVTY MELCEAPSLK PKLGLS

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Macromolecule #11: Flagellar radial spoke protein 12

MacromoleculeName: Flagellar radial spoke protein 12 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO / EC number: peptidylprolyl isomerase
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 19.632256 KDa
SequenceString:
MDFESNGAMM EYCKSTGRTY CAFSIQQSSK LLGTVVLELF TDIAPATCAN FIKYIKDGYQ GTPLHRIVPN GWVQGGDIVD GSGKGDPGF VLPDETYSVK HDAPGVLGMA TGGQPHTANT QFYISLSPLP FLDGKRVAFG RVLNKQSLEN LLALQTLPTF Q NERPVPDV VIASCHVIYN PNS

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Macromolecule #12: Flagellar radial spoke protein 14

MacromoleculeName: Flagellar radial spoke protein 14 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 40.597703 KDa
SequenceString: MDSQARIQAN LRHIQAFEAK PNPKALPETA VTRGFEKVAF PKLVRELTCD NAVVRKKSLL AARELLSSPV NHVQCVAAGA TPAIVALLQ DQTDDETRYY AAGTLKLLAA KEVGARDLAQ HSGLDALAAA LEDPSEGVRD EAYGALIEAA RFDSTRRALE A CGSGAVLP ...String:
MDSQARIQAN LRHIQAFEAK PNPKALPETA VTRGFEKVAF PKLVRELTCD NAVVRKKSLL AARELLSSPV NHVQCVAAGA TPAIVALLQ DQTDDETRYY AAGTLKLLAA KEVGARDLAQ HSGLDALAAA LEDPSEGVRD EAYGALIEAA RFDSTRRALE A CGSGAVLP RLMELALLEA QGGAAGRAQQ GLVLLFTCTQ ARHNAGILSQ LVDVAQAIPH LAGLLKPELP MPVRHAAAEL LG ALATRED AKIQAVQVGA VPLLLLAASP SVPVPFATSA VAALGAITIR REGKYAALES PGGLAGLVSV LDPCHEQLCI NAM TAVSNV AEAPEARAIL VASGAGPKLQ HIFETATVEV VKRAAAQAIR QCRFKHLPYE VLPGAPPINE E

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Macromolecule #13: Flagellar radial spoke protein 16

MacromoleculeName: Flagellar radial spoke protein 16 / type: protein_or_peptide / ID: 13 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 39.052676 KDa
SequenceString: MTRGLDYYEV MGLTRSANDI DIRRAYRRLA LKYHPDINKD GAAGDEFLRI CEAYEVLCDP KTKGFYDLYG EDALKDGISD GNGGLKGPM YRFNPEESPK AVFERFFGTA NPYEALEALS NQFESMTSEE APARGKNKVY PLELTLEEIF HGCLKKVAHK R KVLLFSGE ...String:
MTRGLDYYEV MGLTRSANDI DIRRAYRRLA LKYHPDINKD GAAGDEFLRI CEAYEVLCDP KTKGFYDLYG EDALKDGISD GNGGLKGPM YRFNPEESPK AVFERFFGTA NPYEALEALS NQFESMTSEE APARGKNKVY PLELTLEEIF HGCLKKVAHK R KVLLFSGE YVEEERQLTV DVKPGLPTGT RFVFEGEGNK TPKKEPGPVV FVLKPKPHPR FVRRGSDLVH KVTLPLHHAL IG TTLDIRT LDDRDLKVPI SDIMRPGSSL TVPGEGMPLP ATPSARGNLV IEIDLLFPTH LTETQKMLLR SAFFLPPPTE QNE ETKKAL RDYEAAFKHD LKGWATVFKR

+
Macromolecule #14: Dynein 8 kDa light chain, flagellar outer arm

MacromoleculeName: Dynein 8 kDa light chain, flagellar outer arm / type: protein_or_peptide / ID: 14 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 10.336775 KDa
SequenceString:
MASGSSKAVI KNADMSEEMQ ADAVDCATQA LEKYNIEKDI AAYIKKEFDR KHNPTWHCIV GRNFGSYVTH ETKHFIYFYL GQVAILLFK SG

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Macromolecule #15: Unknown protein

MacromoleculeName: Unknown protein / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 2.91158 KDa
SequenceString:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)

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Macromolecule #16: Flagellar radial spoke protein 23

MacromoleculeName: Flagellar radial spoke protein 23 / type: protein_or_peptide / ID: 16 / Number of copies: 2 / Enantiomer: LEVO / EC number: nucleoside-diphosphate kinase
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 61.422211 KDa
SequenceString: MAELEKTFAL IKPDAVRAGK AQEIMQLIEL NGFTIIAKQK LQLTRARAEE FYGEHKGKEF FPKLVNFMTS GPIWALVLAK PGAILAWRA LMGPTNVFKA RAEQPKCLRA LYGTDGTQNA THGSDSPISA AREIKFFFPT LSGDPTIYAE PTAAAEYITK R IQPALAKA ...String:
MAELEKTFAL IKPDAVRAGK AQEIMQLIEL NGFTIIAKQK LQLTRARAEE FYGEHKGKEF FPKLVNFMTS GPIWALVLAK PGAILAWRA LMGPTNVFKA RAEQPKCLRA LYGTDGTQNA THGSDSPISA AREIKFFFPT LSGDPTIYAE PTAAAEYITK R IQPALAKA LAALAREKPS ADKFEAITFV AGYLLQNNPN KPKVLMPDEW DPALMGDDED DEADFINARL AAPTGNDGAT KA EFDAMVE AAKADTGGAA PAQEFVYPDS KPTTPVVPAP PAPGSKPPSA SGARPQSARP TSARPPSASA APPAPLAPVP PPA SSSRPA SGSGRPPSAT ARPPSATPPP PPPAVELEEA DDPAQLDEAA TKVQAAFRGY QARKEVAVMR SEAQPGEAAA EPEQ EAELQ PEAEPEPQPE GEQEPQPQAS ASSSFLPDGV TEEMAAEAAT RVQAHMRGHL ARKQVAAIKA QQAAPAVAES SEALA EPEP QPEAEAEPQP QASVSSSFLP DGVTEEMAAE AATLVQAHMR GHLARKQVAA IKAQQAAPAV AESSEAQAEA EQTEAE AEP QPDAEAEAGA AEGEAEPEPE AAE

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Macromolecule #17: Cytochrome b5 heme-binding domain-containing protein

MacromoleculeName: Cytochrome b5 heme-binding domain-containing protein / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 26.60907 KDa
SequenceString: MAPPRGPLRR YYTPYEVAMH NTPDDCWVSF LGGVYNLTDL IKANQGALAA PLIAAAGQDL THWFDPTTKD PKRHICPATH IERFYTPMG RFIHVPPPEP MCNWDTSFGL PWWRDAKKYQ IGLLSARTRV IRVRNVLTDQ EDQIEVPCEE KLVEIRERYL E LNAHAKSY ...String:
MAPPRGPLRR YYTPYEVAMH NTPDDCWVSF LGGVYNLTDL IKANQGALAA PLIAAAGQDL THWFDPTTKD PKRHICPATH IERFYTPMG RFIHVPPPEP MCNWDTSFGL PWWRDAKKYQ IGLLSARTRV IRVRNVLTDQ EDQIEVPCEE KLVEIRERYL E LNAHAKSY TWKALVRDPN GDTHVFQELD LNLTLEENGV PDETPVFEDH HVPTDYFIPV LHVYWNDDLT VA

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Macromolecule #18: Uncharacterized protein

MacromoleculeName: Uncharacterized protein / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 75.037945 KDa
SequenceString: MSDPEAEQGE QGYEESPEEP GPGSEAPSPS RIDNGLDTII DIDPQTQHAE EGSNTAYESE QPDVISSYTG GQQEEDGEQA GNGAIDETT EEAAGEADDG GKASGFAVEV DAGTDAAAEG DLEPEPEPER PASASGEPQP TASTSRPASG AAARPASARP T SARPGSAA ...String:
MSDPEAEQGE QGYEESPEEP GPGSEAPSPS RIDNGLDTII DIDPQTQHAE EGSNTAYESE QPDVISSYTG GQQEEDGEQA GNGAIDETT EEAAGEADDG GKASGFAVEV DAGTDAAAEG DLEPEPEPER PASASGEPQP TASTSRPASG AAARPASARP T SARPGSAA PRQPSASGGS RPGSGHPVNL APDSVGLAQQ QQQKSQIEVG AQAYEARGSS RPQSGGDAYG QAEEASAAAA AG RPSTSQS GSRPPPSREG VAVVPSIPED QPLAVPIHIE RYIAPGLKAI EVEVAQGPGM PHRLVRVLLD YTQCDAKPYL GGF RNKRTG AVYHHGATQT PRAPKYSEAD RKLSRETQTV KIKQHSQQTV REQATQMARP GVLLDNDYDK EVTPGRYQTA DERD EIVLR STLRIQRWVR GWLGRKRAAY LRGKKMEREA FLRDQEARAQ SEAEEHRRRE IQRRMHPRTA ADFEVLYNEL EAWRL QETR KIKEAGLAKE QEQQVLQQLL HKETKLLQTI DRLKINANQE NKEARIQHTL NEMSKPKKFA LRNGGKVDVH TPFTTR AKE LQQLYNGLNL PLLTVDERLD VLLHVKWTVK EFDCDLTREL VDLIDREADL LNRGRNPKML EGLRKRISSL FLNFIET PE FNPEAVRFQI VPMDFEAYLY EQVGKATAKA GTSVGTRTLS

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Macromolecule #19: Predicted protein

MacromoleculeName: Predicted protein / type: protein_or_peptide / ID: 19 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 8.111163 KDa
SequenceString:
MSSQYSESRP LHGSAAQTPL YEKMRDYAHS NKVQELLQKM TADLFIERPA DPVTWMIKWL SEEQKRSGTA

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Macromolecule #20: PHOSPHATE ION

MacromoleculeName: PHOSPHATE ION / type: ligand / ID: 20 / Number of copies: 4 / Formula: PO4
Molecular weightTheoretical: 94.971 Da
Chemical component information

ChemComp-PO4:
PHOSPHATE ION / Phosphate

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.75 mg/mL
BufferpH: 7.4
GridPretreatment - Type: GLOW DISCHARGE / Details: unspecified
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV
DetailsThis sample was monodisperse.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 60.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 2320543
CTF correctionSoftware: (Name: CTFFIND, RELION)
Startup modelType of model: OTHER
Details: Negative staining map calculated using the same sample
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 221836

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