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- EMDB-41680: Characterization of the Chlamydomonas Flagellar Mastigoneme Filam... -

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Basic information

Entry
Database: EMDB / ID: EMD-41680
TitleCharacterization of the Chlamydomonas Flagellar Mastigoneme Filament Subunit MST1 Structure at 3.9 angstrom
Map data
Sample
  • Organelle or cellular component: Mastigoneme
    • Protein or peptide: Mastigoneme-like protein
KeywordsFibrous Flagellar Hairs / STRUCTURAL PROTEIN
Function / homologyTyrosine-protein kinase ephrin type A/B receptor-like / Tyrosine-protein kinase ephrin type A/B receptor-like / Putative ephrin-receptor like / Growth factor receptor cysteine-rich domain superfamily / Mastigoneme-like protein
Function and homology information
Biological speciesChlamydomonas reinhardtii (plant)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.9 Å
AuthorsYue W / Kai Z
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35GM142959 United States
CitationJournal: J Cell Biol / Year: 2023
Title: Cryo-EM reveals how the mastigoneme assembles and responds to environmental signal changes.
Authors: Yue Wang / Jun Yang / Fangheng Hu / Yuchen Yang / Kaiyao Huang / Kai Zhang /
Abstract: Mastigonemes are thread-like structures adorning the flagella of protists. In Chlamydomonas reinhardtii, filamentous mastigonemes find their roots in the flagella's distal region, associated with the ...Mastigonemes are thread-like structures adorning the flagella of protists. In Chlamydomonas reinhardtii, filamentous mastigonemes find their roots in the flagella's distal region, associated with the channel protein PKD2, implying their potential contribution to external signal sensing and flagellar motility control. Here, we present the single-particle cryo-electron microscopy structure of the mastigoneme at 3.4 Å. The filament unit, MST1, consists of nine immunoglobulin-like domains and six Sushi domains, trailed by an elastic polyproline-II helix. Our structure demonstrates that MST1 subunits are periodically assembled to form a centrosymmetric, non-polar filament. Intriguingly, numerous clustered disulfide bonds within a ladder-like spiral configuration underscore structural resilience. While defects in the mastigoneme structure did not noticeably affect general attributes of cell swimming, they did impact specific swimming properties, particularly under varied environmental conditions such as redox shifts and heightened viscosity. Our findings illuminate the potential role of mastigonemes in flagellar motility and suggest their involvement in diverse environmental responses.
History
DepositionAug 23, 2023-
Header (metadata) releaseOct 25, 2023-
Map releaseOct 25, 2023-
UpdateNov 8, 2023-
Current statusNov 8, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_41680.png

Downloads & links

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Map

FileDownload / File: emd_41680.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.149 Å
Density
Contour LevelBy AUTHOR: 0.0728
Minimum - Maximum-0.222222 - 0.78560156
Average (Standard dev.)0.007356265 (±0.025676329)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin4948
Dimensions512512512
Spacing512512512
CellA=B=C: 588.288 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_41680_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_41680_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Mastigoneme

EntireName: Mastigoneme
Components
  • Organelle or cellular component: Mastigoneme
    • Protein or peptide: Mastigoneme-like protein

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Supramolecule #1: Mastigoneme

SupramoleculeName: Mastigoneme / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all
Details: Fibrous Flagellar Hairs of Chlamydomonas reinhardtii
Source (natural)Organism: Chlamydomonas reinhardtii (plant)

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Macromolecule #1: Mastigoneme-like protein

MacromoleculeName: Mastigoneme-like protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Chlamydomonas reinhardtii (plant)
Molecular weightTheoretical: 205.404203 KDa
Recombinant expressionOrganism: Chlamydomonas reinhardtii (plant)
SequenceString: MAMPRHGRRP ARCSSNRASQ WLLLVLGVAL AASPRFLLLV EAATYTLTLD KLGPTVNPTT SDAVTFTATV ASPDSTTAVF FTLDYGDGV TAETTRTTTG ALSTTPTANL VSAGTYTVTY ASIGTKFVTL RLYDSAVAPG VLLASKTVPI YVEDSTLTAT L LQSGVPRL ...String:
MAMPRHGRRP ARCSSNRASQ WLLLVLGVAL AASPRFLLLV EAATYTLTLD KLGPTVNPTT SDAVTFTATV ASPDSTTAVF FTLDYGDGV TAETTRTTTG ALSTTPTANL VSAGTYTVTY ASIGTKFVTL RLYDSAVAPG VLLASKTVPI YVEDSTLTAT L LQSGVPRL NLAFSGFKGR VSSSTANRAD MWATIQLDTA PGVFESSRIF IGIAPTASTN YDFVIPDQVY NLEGAKTTVL RI YDAPVGG TLLRTFTPAA ANAVYVVDPS KYVLTLTVGP TSVTTADQVT FTQTTTEVSY SASATSPILQ WRFNWDDPSV VET PLAYPD ALTAASNFPT TATAVSSAAA STFRYTSTGS KNARLRLYDG ANNVIAEKIV VITVSNAGYT LALAKTTADP VTTD DTIAF SAGAKHLSST SQVWWTIDYG AGESSPRTAL TMTNVGAAAP NAIASLSNQY TSGGTKLATL RIYDRDGVGA NTGLL LAST TVTFTVTPVL YALESAVEPF SPIATVAAKW SFRIQRSKAT PAGVTESIKC AFFGADTGTA PADLAAWLTA ANGAGG LTA TILPSSIPSD IISFTRTYAA AAASLQGKLQ CFIGSTPLWD PYYPTPVFQV LAAAPTYTLS ASVTPAVVPV DTATLWT YN IIRSVPVPAG GPSLPILCSF WDGKTGAAPT TDAGWAALAG SANGKGTSMA PGSTTATCSF TPSYSTTGTA TPTLQLIQ N SFALDAATTV GFLSPVYTAP AFATVTAASY TISSYLNPVT PVAGGAAAVW RIVITRNAAV TASAKTLTCQ MPDNGQGGS PADVTADIAV GGTTTVCVFS IAGYTTATPG PYFATVNVVD GAVTTSHITK NFTVLASGTT APTYAVTSVV SPATPVKVST PVTYTFTIT RTTAVPAGGI PQPIICEFFN GEGTAPASAA AYWRVSTTIP DADTVVAVMA PGETTTTCTF TTYYTTVSAG G FTAKLMVF GESATAAPLL TSLSVTPSQL LAAVHSFATP MVVAAAVVAV ESTTISPNYN PTTPYTNIPT YFTFTLLRDP PV PPSASSG VQFACALYTG QNVNPASAPS AITDAVYKTF TDVTTAVATD ANYFADQQLR VVTMAPGTGR VSCTFPTLYA AAG PFSPKF FVFEYASSTV GANALAVADT VTSLTSFTTQ AAPTFITGPT NVPQRVPLPK GFRTTCFDGY ELIFSNDNYT NGVR VAVDA YPYPVGQCRK CPGGTATMDG YRCIPCPSGY WSNEGARECT ACPAGTIAKP AALTARAKYS IDPTTYHFVT HLAMG PESC KKCPKGYFQP NIAGTVCLPC PSGFVSTSGA TGCTACSEGT YHTDGVGTTT PGEATSLDTT DTFGSIYPII PNTCRQ CPA NTYLPLRGQA AIASMNLAAV SSATPCRPCE DGTWSKAGAA GCQKCPPGTY RNTWFSGQLG SPFITADGVP VATTLTE LG SGCSQCPPGT YAPTFGMSVC LPCPAGTFAS APGATACQQC KPGTNSLMGD RTQQMALVVT NAANDFPALR AYTISGMV A GPAYAKPIVT GPDTNFFMAG KSETCSTNLP GYYTDVDGLP IQLPCKPGTF MPFDTATANL LDTGLTVDGT QCYTCQTGT FNDEFSQPVC KACWSGSFAS KRGLPTCEIA QPGTFTNVAA AANATFNTAT LIPTGLVKGA QAPTPCGMGY FQSSAETTTC TACAVGTYA DQAGLAACKP CQPGRYQNSI GQRVCKPCDM GTYSRYGGEL CTKCPAGTVA SKTGSSQCTP CAAGFYANAP D SATSCRAC PRGYYGPYSG AYADNLGDEF EGPRGCYKCP YDFFADRPGV RQCTACPPLD LGGGNLVEQC TEDLGSQRCK PC SLLSKPK TARTEQSPPP PSPSPPPPPP PSPRPPSPNP PSPRPPSPAP PSPNPPPTSP PPSPPPSPPP PRPPPPPPPP PSP PPPNRS PPPPPPASSA INPGGGVNQN GDPVGHRRAI LSLMEDEDAE AAQEEQAIVD VDAEMQPQDD E

UniProtKB: Mastigoneme-like protein

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation statefilament

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Sample preparation

BufferpH: 7.4
Details: 30 mM HEPES, 5 mM MgSO4, 1 mM DTT, 0.5 mM EGTA, 25 mM KCl, 1 mM PMSF, pH 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS GLACIOS
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.5 µm / Nominal defocus min: -0.9 µm
Image recordingFilm or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 40.0 e/Å2

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: NOT APPLICABLE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 70074
FSC plot (resolution estimation)

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