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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-22444 | ||||||||||||
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Title | Chlamydomonas reinhardtii radial spoke minimal head complex | ||||||||||||
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Function / homology | ![]() radial spoke head / radial spoke / kinocilium / cilium movement involved in cell motility / motile cilium assembly / axoneme assembly / motile cilium / axoneme / membrane Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.95 Å | ||||||||||||
![]() | Grossman-Haham I / Coudray N / Yu Z / Wang F / Bhabha G / Vale RD | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure of the radial spoke head and insights into its role in mechanoregulation of ciliary beating. Authors: Iris Grossman-Haham / Nicolas Coudray / Zanlin Yu / Feng Wang / Nan Zhang / Gira Bhabha / Ronald D Vale / ![]() Abstract: Motile cilia power cell locomotion and drive extracellular fluid flow by propagating bending waves from their base to tip. The coordinated bending of cilia requires mechanoregulation by the radial ...Motile cilia power cell locomotion and drive extracellular fluid flow by propagating bending waves from their base to tip. The coordinated bending of cilia requires mechanoregulation by the radial spoke (RS) protein complexes and the microtubule central pair (CP). Despite their importance for ciliary motility across eukaryotes, the molecular function of the RSs is unknown. Here, we reconstituted the Chlamydomonas reinhardtii RS head that abuts the CP and determined its structure using single-particle cryo-EM to 3.1-Å resolution, revealing a flat, negatively charged surface supported by a rigid core of tightly intertwined proteins. Mutations in this core, corresponding to those involved in human ciliopathies, compromised the stability of the recombinant complex, providing a molecular basis for disease. Partially reversing the negative charge on the RS surface impaired motility in C. reinhardtii. We propose that the RS-head architecture is well-suited for mechanoregulation of ciliary beating through physical collisions with the CP. | ||||||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 117.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 18.9 KB 18.9 KB | Display Display | ![]() |
Images | ![]() | 72.9 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7jr9MC ![]() 7jrjC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 0.855 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : radial spoke minimal head complex
Entire | Name: radial spoke minimal head complex |
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Components |
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-Supramolecule #1: radial spoke minimal head complex
Supramolecule | Name: radial spoke minimal head complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Macromolecule #1: Radial spoke protein 9
Macromolecule | Name: Radial spoke protein 9 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 29.572348 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: 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 |
-Macromolecule #2: Flagellar radial spoke protein 4
Macromolecule | Name: Flagellar radial spoke protein 4 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 52.159195 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGSSHHHHHH GGSAENLYFQ GMAAVDSVAQ ALAYLQVHSP QDGTSMYDHL VKLVSKVLED QPKNAVDLLE TSLLVKKSTF DPKESSPLV PIPVAPDATQ TQAAVSIFGD PELPINPATG EPVPADPPNE FEAENMLGAA AVLDCLGVGL GRELGVNIAL A AKRIGEDP ...String: MGSSHHHHHH GGSAENLYFQ GMAAVDSVAQ ALAYLQVHSP QDGTSMYDHL VKLVSKVLED QPKNAVDLLE TSLLVKKSTF DPKESSPLV PIPVAPDATQ TQAAVSIFGD PELPINPATG EPVPADPPNE FEAENMLGAA AVLDCLGVGL GRELGVNIAL A AKRIGEDP KLAVRSVRFF GKFLGLYSDY FVFEVAFKKE AAKEAAPAAP APERVEGEAA SSSAPEVPVE EPGKGANKFT YL VCSSLGG PLTRLPDVTP AQVKASRRIK KLLTGRLTSH VSTYPAFPGN EANYLRALIA RISAATVVAP SDLFSLNDET GEL ERAEDW EPPAGREMAA PTAWVHVRPH LKSQGRCEVH KRELPEDADE DEFYNEDELE EGPDLLAALE EDAQLPGEQA AWTP IYSSA SEAVKTQAGG LRSLVWPGAV CGGRGSEWTC VYVGWGVKNA PFVPLPPPPV AQEFAWGEVE TQELELKPAP PPPEE EAEA DE |
-Macromolecule #3: Flagellar radial spoke protein 6
Macromolecule | Name: Flagellar radial spoke protein 6 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 48.884551 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: 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 |
-Macromolecule #4: Flagellar radial spoke protein 6
Macromolecule | Name: Flagellar radial spoke protein 6 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 698.854 Da |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) |
-Macromolecule #5: Flagellar radial spoke protein 10
Macromolecule | Name: Flagellar radial spoke protein 10 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 528.644 Da |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) |
-Macromolecule #6: Radial spoke protein 10
Macromolecule | Name: Radial spoke protein 10 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 23.553943 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: 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 |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.1 mg/mL |
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Buffer | pH: 7.4 / Details: 30 mM HEPES, 150 mM NaCl, 1 mM TCEP |
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GRAPHENE OXIDE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Slit width: 30 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number grids imaged: 2 / Number real images: 2886 / Average electron dose: 73.3 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.001 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 130000 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Protocol: AB INITIO MODEL |
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Output model | ![]() PDB-7jr9: |