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- EMDB-64028: The overall cryo-EM map of the mouse CatSper trimer -

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Basic information

Entry
Database: EMDB / ID: EMD-64028
TitleThe overall cryo-EM map of the mouse CatSper trimer
Map data
Sample
  • Complex: Homotrimer of the mouse CatSper
    • Protein or peptide: x 14 types
  • Ligand: x 2 types
Keywordssperm / hyperactivation / calcium channel / PROTEIN TRANSPORT
Function / homology
Function and homology information


Sperm Motility And Taxes / CatSper complex / fusion of sperm to egg plasma membrane involved in single fertilization / sperm principal piece / regulation of cilium beat frequency involved in ciliary motility / calcium ion sensor activity / male meiotic nuclear division / organic anion transport / flagellated sperm motility / : ...Sperm Motility And Taxes / CatSper complex / fusion of sperm to egg plasma membrane involved in single fertilization / sperm principal piece / regulation of cilium beat frequency involved in ciliary motility / calcium ion sensor activity / male meiotic nuclear division / organic anion transport / flagellated sperm motility / : / calcium-activated cation channel activity / fertilization / motile cilium / sodium ion transport / sperm capacitation / regulation of calcium ion transport / monoatomic ion channel complex / sperm flagellum / voltage-gated calcium channel activity / acrosomal vesicle / establishment of localization in cell / calcium ion transport / spermatogenesis / cell differentiation / calmodulin binding / cilium / calcium ion binding / endoplasmic reticulum / plasma membrane / cytoplasm
Similarity search - Function
Cation channel sperm-associated auxiliary subunit TMEM249 / Cation channel sperm-associated auxiliary subunit gamma / Cation channel sperm-associated protein 4 / Cation channel sperm-associated protein 1 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated auxiliary subunit beta / Cation channel sperm-associated auxiliary subunit delta/epsilon / Cation channel sperm-associated auxiliary subunit TMEM262 / : / : ...Cation channel sperm-associated auxiliary subunit TMEM249 / Cation channel sperm-associated auxiliary subunit gamma / Cation channel sperm-associated protein 4 / Cation channel sperm-associated protein 1 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated auxiliary subunit beta / Cation channel sperm-associated auxiliary subunit delta/epsilon / Cation channel sperm-associated auxiliary subunit TMEM262 / : / : / : / : / : / : / : / : / : / : / : / : / : / : / : / : / CATSPERD, beta-propeller domain / CATSPERG beta-propeller domain / CATSPERB, C-terminal domain / Cation channel sperm-associated auxiliary subunit TMEM249 / CATSPER protein subunit beta protein 1st domain / CATSPER subunit beta protein 2nd domain / CATSPERB head domain / CATSPERB Ig-like domain / CATSPERG N-terminal domain / CATSPERE first N-terminal domain / CATSPERE second N-terminal domain / CATSPERE beta-propeller domain / CATSPERG C-terminal domain / CATSPERE Ig-like domain / CATSPERD/E C-terminal domain / CATSPERG Ig-like domain / CATSPERD Ig-like domain / Cation channel sperm-associated auxiliary subunit zeta / EF-hand calcium-binding domain-containing protein 9 / Organic anion transporter polypeptide / Organic Anion Transporter Polypeptide (OATP) family / Kazal-type serine protease inhibitor domain / Kazal domain superfamily / Kazal domain / Kazal domain profile. / Voltage-dependent channel domain superfamily / MFS transporter superfamily / EF-hand calcium-binding domain profile. / EF-hand domain / Ion transport domain / Ion transport protein / EF-hand domain pair
Similarity search - Domain/homology
Cation channel sperm-associated auxiliary subunit TMEM249 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated auxiliary subunit beta / Cation channel sperm-associated auxiliary subunit gamma 2 / Cation channel sperm-associated auxiliary subunit TMEM262 / Cation channel sperm-associated auxiliary subunit delta / Cation channel sperm-associated auxiliary subunit epsilon / Cation channel sperm-associated protein 3 / Cation channel sperm-associated protein 4 / Solute carrier organic anion transporter family member 6C1 ...Cation channel sperm-associated auxiliary subunit TMEM249 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated auxiliary subunit beta / Cation channel sperm-associated auxiliary subunit gamma 2 / Cation channel sperm-associated auxiliary subunit TMEM262 / Cation channel sperm-associated auxiliary subunit delta / Cation channel sperm-associated auxiliary subunit epsilon / Cation channel sperm-associated protein 3 / Cation channel sperm-associated protein 4 / Solute carrier organic anion transporter family member 6C1 / Cation channel sperm-associated protein 1 / Cation channel sperm-associated auxiliary subunit zeta / EF-hand calcium-binding domain-containing protein 9
Similarity search - Component
Biological speciesMus musculus (house mouse)
Methodsingle particle reconstruction / cryo EM / Resolution: 6.8 Å
AuthorsXu Q / Lin S / Zhao Q / Yan Z / Wu J
Funding support China, 1 items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC) China
CitationJournal: To Be Published
Title: Cryo-EM structure of the mouse CatSper trimer
Authors: Xu Q / Lin S / Zhao Q / Yan Z / Wu J
History
DepositionApr 3, 2025-
Header (metadata) releaseDec 31, 2025-
Map releaseDec 31, 2025-
UpdateDec 31, 2025-
Current statusDec 31, 2025Processing site: PDBc / Status: Released

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Structure visualization

Supplemental images

emd_64028.png

Downloads & links

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Map

FileDownload / File: emd_64028.map.gz / Format: CCP4 / Size: 824 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.09 Å/pix.
x 600 pix.
= 652.2 Å		1.09 Å/pix.
x 600 pix.
= 652.2 Å		1.09 Å/pix.
x 600 pix.
= 652.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.087 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.245
Minimum - Maximum-0.37576637 - 0.9998739
Average (Standard dev.)0.001660244 (±0.046525996)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions600600600
Spacing600600600
CellA=B=C: 652.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_64028_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_64028_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 : Homotrimer of the mouse CatSper

EntireName: Homotrimer of the mouse CatSper
Components
  • Complex: Homotrimer of the mouse CatSper
    • Protein or peptide: Cation channel sperm-associated protein 1
    • Protein or peptide: Cation channel sperm-associated protein 2
    • Protein or peptide: Cation channel sperm-associated protein 3
    • Protein or peptide: Cation channel sperm-associated protein 4
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit beta
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit gamma 2
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit delta
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit epsilon
    • Protein or peptide: EF-hand calcium-binding domain-containing protein 9
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit TMEM249
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit zeta
    • Protein or peptide: Solute carrier organic anion transporter family member 6C1
    • Protein or peptide: Cation channel sperm-associated auxiliary subunit TMEM262
    • Protein or peptide: Unknown protein
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
  • Ligand: CHOLESTEROL

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Supramolecule #1: Homotrimer of the mouse CatSper

SupramoleculeName: Homotrimer of the mouse CatSper / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#14
Source (natural)Organism: Mus musculus (house mouse)

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Macromolecule #1: Cation channel sperm-associated protein 1

MacromoleculeName: Cation channel sperm-associated protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 29.080652 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: LLGLQQMILS LTQSLGFETF IFIVVCLNTV ILVAQTFTEL EIRGEWYFMV LDSIFLSIYV LEAVLKLIAL GLEYFYDPWN NLDFFIMVM AVLDFVLLQI NSLSYSFYNH SLFRILKVFK SMRALRAIRV LRRLSILTSL HEVAGTLSGS LPSITAILTL M FTCLFLFS ...String:
LLGLQQMILS LTQSLGFETF IFIVVCLNTV ILVAQTFTEL EIRGEWYFMV LDSIFLSIYV LEAVLKLIAL GLEYFYDPWN NLDFFIMVM AVLDFVLLQI NSLSYSFYNH SLFRILKVFK SMRALRAIRV LRRLSILTSL HEVAGTLSGS LPSITAILTL M FTCLFLFS VVLRALFQDS DPKRFQNIFT TLFTLFTMLT LDDWSLIYID NRAQGAWYII PILMIYIVIQ YFIFLNLVIA VL VDNFQMA LLKG

UniProtKB: Cation channel sperm-associated protein 1

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Macromolecule #2: Cation channel sperm-associated protein 2

MacromoleculeName: Cation channel sperm-associated protein 2 / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 33.115477 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: PLSLWAGWVL DSSVFSKFII SLIFLNTFVL MVEIELMEST NTALWPVKLA LEVADWFILL SFIVEILLMW LASFSLFWKD AWNVFDFFV TLLSLLPELV VLLGVPAHSV WLQLLRVCRV LRSLKLFARF RQIKVILLAL VRALKSMTFL LMLLLIFFYI F AVTGVYFF ...String:
PLSLWAGWVL DSSVFSKFII SLIFLNTFVL MVEIELMEST NTALWPVKLA LEVADWFILL SFIVEILLMW LASFSLFWKD AWNVFDFFV TLLSLLPELV VLLGVPAHSV WLQLLRVCRV LRSLKLFARF RQIKVILLAL VRALKSMTFL LMLLLIFFYI F AVTGVYFF REYSRSTIEG LEYNMFFSDL LNSLVTVFIL FTLDHWYAVL QDIWKVPESS RVFSSIYVIL WLLLGSIIFR NI IVAMMVT NFQNIRSELS EEMSHLEVQY KADMFKQQII QRRQ

UniProtKB: Cation channel sperm-associated protein 2

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Macromolecule #3: Cation channel sperm-associated protein 3

MacromoleculeName: Cation channel sperm-associated protein 3 / type: protein_or_peptide / ID: 3 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 32.308949 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: ECQAYFRKVI KSTFFQIVMI TTVTTNSFLL VLGTNYDIQF EFFRTFEVSE LFFVSVYVCE FLMKVYVDPI TYWKDGYNIL DVIILIILT IPYLLRKIKG NHSAYLHFAD GIQSLRILKL ISYSRGIRTL IIAVGETVYT VASVLTLLFL LMFVFAILGF C LFGVTDRG ...String:
ECQAYFRKVI KSTFFQIVMI TTVTTNSFLL VLGTNYDIQF EFFRTFEVSE LFFVSVYVCE FLMKVYVDPI TYWKDGYNIL DVIILIILT IPYLLRKIKG NHSAYLHFAD GIQSLRILKL ISYSRGIRTL IIAVGETVYT VASVLTLLFL LMFVFAILGF C LFGVTDRG DLENWGNLAS AFFTLFSLAT VDGWTDLQEE LDKRKFTVSR AFTILFILLA SFIFLNMFVG VMIMHTEDSM KK FERDLTL ERNLAIMEEK QIILKRQQEE VNRLMNTQKS G

UniProtKB: Cation channel sperm-associated protein 3

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Macromolecule #4: Cation channel sperm-associated protein 4

MacromoleculeName: Cation channel sperm-associated protein 4 / type: protein_or_peptide / ID: 4 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 28.88043 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: DVMEKKDAWD VQEFITQMYI KQLLRHPAFQ LLLAFLLLSN AITIALRTNS YLGQKHYELF STIDDIVLTI LICEVLLGWL NGFWIFWKD GWNILNFAIV FILFMGFFIK QLDMVAITYP LRVLRLVHVC MAVEPLARII KVILQSMPDL ANVMALILFF M LVFSVFGV ...String:
DVMEKKDAWD VQEFITQMYI KQLLRHPAFQ LLLAFLLLSN AITIALRTNS YLGQKHYELF STIDDIVLTI LICEVLLGWL NGFWIFWKD GWNILNFAIV FILFMGFFIK QLDMVAITYP LRVLRLVHVC MAVEPLARII KVILQSMPDL ANVMALILFF M LVFSVFGV TLFGAFVPKH FQNMGVALYT LFICITQDGW LDIYTDFQMD EREYAMEVGG AIYFAVFITL GAFIGLNLFV VV VTTNLEQ MMK

UniProtKB: Cation channel sperm-associated protein 4

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Macromolecule #5: Cation channel sperm-associated auxiliary subunit beta

MacromoleculeName: Cation channel sperm-associated auxiliary subunit beta
type: protein_or_peptide / ID: 5 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 121.580594 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: VIHNKGKERT YFSCSGEGIL TGLHTIKLFL TMDNLKVRCF FRNENQSPSK EILGLFTSGG LAPNMIITNS TFYGGYYFKL TPFSNRLEW LIDIPRQNIT VNTDIAAVEQ WMIKITMHEG LNIYDTEGTL LDLVREPILQ WNLGRVLTEM EVRDLYPEVN D IKVTKSPC ...String:
VIHNKGKERT YFSCSGEGIL TGLHTIKLFL TMDNLKVRCF FRNENQSPSK EILGLFTSGG LAPNMIITNS TFYGGYYFKL TPFSNRLEW LIDIPRQNIT VNTDIAAVEQ WMIKITMHEG LNIYDTEGTL LDLVREPILQ WNLGRVLTEM EVRDLYPEVN D IKVTKSPC ANDVALIGFM MKPSSNGVFI GKTISGFWTY KECIWHDLTE IIYAELKDEH QGLTVIDLVL TNHFLVILTS LG LYVSSDL RYPTTSQIKL SRAEFCGFER VDYIRGNLWY NEKCFANRES FEVDYVTITF NRNRTLSESS SCFFSKEPFL HWL PCVFST IKNEKSIPRV ITFLIDQETD SGIYLFNVQD TKETYVTVAM LKDGKPSPRP KFPSFHFPST FTLPLGMIFH PRSH FLYVY GSQIWVSMDG GNTFEMLCNL FSHHVTKTSN SFYTSDIVFI VEDGRILTTK AGLTTYSELG ILKDAIFTLY YDQLG YIHK LTPENFDAGS KLLGHGNSGS IFGKRPDLGF EAILVPQYIS TNEMYFFAHV PLTMPTNIQW KKRFKTIHLG KTIEFS KTG LANIKNVYMH KTEPVGFQTS IHTEIIVPFG IENSKDSPCL LSDLEITYSG KLYYTIKLLS KNPLHELKST DVEKSVL IP GYSSFLIMNI TDKWTASALA TMPQAIKSNL KFLTGSWFLY NFGTAGGRKW SISTRQCNYW IQQDSLDFMS LNLVKYID V GNTIDFQFKI IPKAMSTFPI PPVSMVVGNP GLVEVKTQGV FDLNENYYLD IHVSGRFFQK GSTSIALVLW EGSSKCYAI TLLPTIKSSC SYLRTMHHTP GRHIPPEDWI SGVHKDSQGF NMIKTLPINY RPPSHMGISI PLTDNFYHAD PSKPIPRNQF HKSKETGKY KQCANVTSRA MCNCSEHQKF SHAVAFSDCK EKVHRFKFPV TQYPVVLEIF NERDKISAEP PYLVTMTEVN M RKNWQLKH NEPENVKKMK HYLEPLLKTP VYNPLGLNLT IQGSELFHFK VSVVPGVSFC ELSEEFQIYV DEVPLPFPGH AL IAVATSV VLGVLIFIAF VFQLRNIH

UniProtKB: Cation channel sperm-associated auxiliary subunit beta

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Macromolecule #6: Cation channel sperm-associated auxiliary subunit gamma 2

MacromoleculeName: Cation channel sperm-associated auxiliary subunit gamma 2
type: protein_or_peptide / ID: 6 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 119.864367 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: HCTWLLVLNK FEKVGLHLSK DRFQDHEPID TVAKVFQKLT DSPIDPSENY LSFPYYLQIN FSCPGQNIEE LARKGHLMGM KPMVQINYM YSVNFYRWEM ENVQILMEAA PMRSTGYCPA EAMCVLNWYT PMPFKNGSVV SSVDIYTNGI GPFVSKKRFY V NMNGFLKR ...String:
HCTWLLVLNK FEKVGLHLSK DRFQDHEPID TVAKVFQKLT DSPIDPSENY LSFPYYLQIN FSCPGQNIEE LARKGHLMGM KPMVQINYM YSVNFYRWEM ENVQILMEAA PMRSTGYCPA EAMCVLNWYT PMPFKNGSVV SSVDIYTNGI GPFVSKKRFY V NMNGFLKR DASGKSLFAI GYESLVLKSS HFRLSKSRPL WYTVNHAPVF ILGGFYDEKS ILFSDSNFQD YVLLELSIDS CW VGSFYCP ILGFSATIHD AIATESTLFI RQNQLVYYFT GTYITLFDKS HGSSRWVRVL PSECIKRLCP VYASGNGSEY VLA LTTGKN EGYIHIGTIT DGLVSFEMVP DGWSVCEKLP GKNCSIDWAT YIADERNLLL LVKIDSGQFY LVNFNTEFKT LNIL YKIPE FIPEAKELDF LVLLDTVTYT NTPMTPKGLF FNTLNNMLYI WGNFILQSYN REEFIFLADF PKESTIKYMV NSFKG QMAV VTENEEIWYF LEGGYDVYQV VPSQGWETYH NLQKMQKSSF HSEDESLVSL FFEDGKLFQL VYLFDVGKER LVKRLL PVG TLMEYNLPKP FTVVNQGNYQ AISFTHTCPF KEIHLIDVPK KHHASRTESY VALPPLVSES LGFHNNNTLA VYQGLVY YL LWLHSKYDKP YADPVHDPTW RWWQHKTKDK DYFFYLFSNR LAAEGIYINM NAYQKLYNMS GDYGIPDLFF LDKGNWFT I TVVLLSHQDT FTSSDSQGPT INVDKKLAIA VTIADPECLS VTVTQDVLLN RNAVINKIKV IDKKRCSEQG MIGRNIKKT SMMLKVLGAP GNCIQRTYLG GIIQGFKVVP IFIGCPPGKR LAFDVSYTIM HSEEINKHYF DCVIKDAEMP CFLFRDLFQP FFLVQDLVT GDSGSFLGSY VLKVVGGGRT LNTIRDYTEE EIFRYNSPLD TTNSLIWKTK VERTTEDKKF YIMSHESPGV E WLCLENSP CYDIIPQSIY PPEFFFKLLV SNRGVDNSTY CDYKLTFIVH IHGLPLSSKR TSFIVMVSTS FFIALVVFYI LF CL

UniProtKB: Cation channel sperm-associated auxiliary subunit gamma 2

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Macromolecule #7: Cation channel sperm-associated auxiliary subunit delta

MacromoleculeName: Cation channel sperm-associated auxiliary subunit delta
type: protein_or_peptide / ID: 7 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 82.834281 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: HTLCRVHTVR TGKVFKSNIQ LQGDPLFYAF PNTFVLKNVC KADISVYLGQ KVFLTIDNFE SSLLPLTVPK SLAVGVPSIT SAHFVSGSL VLFVISGKGY SYDYYENTWR KLEGISEPVS HISGDVCCFK GSFCLELSNN LFAYLRGGQI PGTNIYFSDN G GFSFQLMN ...String:
HTLCRVHTVR TGKVFKSNIQ LQGDPLFYAF PNTFVLKNVC KADISVYLGQ KVFLTIDNFE SSLLPLTVPK SLAVGVPSIT SAHFVSGSL VLFVISGKGY SYDYYENTWR KLEGISEPVS HISGDVCCFK GSFCLELSNN LFAYLRGGQI PGTNIYFSDN G GFSFQLMN TDKLSHLTGT LGGIFHLHSM SQVGVLMVEN NLGTFHYMEY PLNHSMGIAF SYKNLLEVIM KPYQRGFMVL WN QKSILVS SNSGQIVEHV RLIDQKIFTD LDVEHANINI YSVASNAYEL AFLVAEDHLY YGSQSYMGTY VIKLPHQPLW STH TSIYFE DIGILQVLTP VADPHFAAYD FDKCTVNVQS SLMDEKLALQ PCNVELLEST MINTMFTIDM NSKLKLSALM IPRK GENPT PLVMVSNPHA LGFKANLNEF GNTFDGNSKY KLDIELKQQH HWGNSDFNFT ASIKRHAISS VTVDIADKTL SCVDL KPLS TLISVGCDMT KKIVVQNKIS ACTMGILNPV QLQKNYTYTI EKEAYDPINH NGEAQDDLIV FYEYKDLGCP RLVYYD KPW KPVVELWKNG IVEEIMNAEY VISEINGLVT YSYSLTAATA NCRSQPQNWS TFESDIENEE PFLWNRENYV SCHEDNK DN PLLWPNVEYQ VLGGQTNNKI IFGQRNGIYT FHLSVVDPYY SYCNLNTIFS VYVHGALPVT KFQPLLTILL MVTTTLLT A WLAYAIPKQL RS

UniProtKB: Cation channel sperm-associated auxiliary subunit delta

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Macromolecule #8: Cation channel sperm-associated auxiliary subunit epsilon

MacromoleculeName: Cation channel sperm-associated auxiliary subunit epsilon
type: protein_or_peptide / ID: 8 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 107.004016 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: LWRYYINSQD YSIFSTRSSI KLEYEGNSFV SWKIPESCKV ENTTSPKTTL HCKRAGIHTI KPIAGNQEVE RHLTVDNSYI CYLWYFTVV DVYYNLSQIV TIWVYDPESA STEELIWTAK KPSLSSRVLT KQMNTLGQRP FIFTVEKRLT YHPGPLTSEG T WVIHLPMS ...String:
LWRYYINSQD YSIFSTRSSI KLEYEGNSFV SWKIPESCKV ENTTSPKTTL HCKRAGIHTI KPIAGNQEVE RHLTVDNSYI CYLWYFTVV DVYYNLSQIV TIWVYDPESA STEELIWTAK KPSLSSRVLT KQMNTLGQRP FIFTVEKRLT YHPGPLTSEG T WVIHLPMS SDDIAKVIRG NKVAFQDCFI ANLYFMLTYP MTIISEPPGY EPLTVPPGSP LMLSWDTCIS TFALLATDQE TF QTNDSFQ TWTRVRAPPG ILSDAQRHSL RDVIIFDQGT LFLVDGTVYL RTEDEFTKLD ESRGISETGI LGFSKRRWCQ IRY LYKLAS KKSILIAWSK TTVYAGYATF RFVTLTDTAK LKDFLKLPQT DTLEVMSVEY LWHPLEAAVL LSHCSVCTTN TRNI RIVIY SAIFQTWTLQ DFELQLPKEA ILEFRFLYSA MPDIIMWDQH HVYYSYKNFT VVGTISTPSG ETNLSSLSQG SKIHQ VLTD RIGNVVVKME NNVMFYIKAD ITEAVILHTW VNTTAKTVVL FDKSFEVCIL YYNENLDEKY QLQTQPYPLI LELQSI NKD LGDWCPYLAF QHNIHSQFYH MDKGESLTIW SQIVYPENRG LYIVVEHYGS SVMTWTQNLE YEIASGFCTK TMITRFF QT TNYELVDNYY QLQKENTGLM LLQFRPSEFS RTCLTAKPVF EIDVGCDSSK YIMVRGFNKS RCQRRDFSYV IDKELLRE S LSDNLKVRYD VAKYGCPLTL ELGQMFQPIV ELYDENGFIK IVDANFILWE IHGRNDYTFN STMEQNGCIN EAQTWDSMI EENPDIPLDD VWGPQNYRPC FSYAIGKPGD LGQPYEILNY SNKNHIKWPM TYAGMYVYRL KILDPNYSFC NLTTIFAIES LGMIPRSSV YLVAALIFVL MLTFISILVL SYFWYLKIYR QF

UniProtKB: Cation channel sperm-associated auxiliary subunit epsilon

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Macromolecule #9: EF-hand calcium-binding domain-containing protein 9

MacromoleculeName: EF-hand calcium-binding domain-containing protein 9 / type: protein_or_peptide / ID: 9 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 20.942268 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString:
MKLTPGCFLW YLYMDKIYCL LSLRNVKALM VYFHLLDVHH RNTLNDVLFF HFLQHVTNLN KSQIGMIFDL LDWTAVGEIG FDQFYVLIC ILLAHQDHLE DHFMYRHSRP VFELLDLDGE MNIGAANFQN YRFLFNIKKQ ELRDLFHDFD ITGDRLLNYK E FKLYTIFC TDKSID

UniProtKB: EF-hand calcium-binding domain-containing protein 9

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Macromolecule #10: Cation channel sperm-associated auxiliary subunit TMEM249

MacromoleculeName: Cation channel sperm-associated auxiliary subunit TMEM249
type: protein_or_peptide / ID: 10 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 19.898578 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString:
MCWVMPPSVF VLEYYLDTLW KGTMLFIICL VFISCNVLRE VKYQETWCFP AYGMVIGLWL MLSSIPQRRL VLNHTRGMYH FSIQGRTVC QGPMHLVYVR LALSSDAYGG RFFQLVLCGH KLEPLVLVQL SERYEQMEFL GRHLARKLNI NYFDYLASSY R HVVRHWPL

UniProtKB: Cation channel sperm-associated auxiliary subunit TMEM249

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Macromolecule #11: Cation channel sperm-associated auxiliary subunit zeta

MacromoleculeName: Cation channel sperm-associated auxiliary subunit zeta
type: protein_or_peptide / ID: 11 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 17.504592 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString:
TANVSVSDVC EDFDEEGKSV RNRIRKYSQT ISIRDSLNLE PEEIQQQARR ELELCHGRSL EHGEDHEESE TSLASSTSES LIFSLWKPH RTYWTEQQNR LPLPLMELME TEVLDILKKA LITYRSTIGR NHFMTKELQG YIEGIRKRRN

UniProtKB: Cation channel sperm-associated auxiliary subunit zeta

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Macromolecule #12: Solute carrier organic anion transporter family member 6C1

MacromoleculeName: Solute carrier organic anion transporter family member 6C1
type: protein_or_peptide / ID: 12 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 68.34968 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: PIVLPFVQRF NNIDGFMTLY VAAVLIHGAL FAVVDMTLNI YQVQFSLTRT EWYLMDFSDY IASFVVAIII AHFGSKGNRT RWIAASCIL MGLESMLFAF PFFTYEIIIP GRQSIELCME ENEKRNIICG NSVPNRSKCI YFHIGGQCIH GIAGMPIYIL G ITFIFDHI ...String:
PIVLPFVQRF NNIDGFMTLY VAAVLIHGAL FAVVDMTLNI YQVQFSLTRT EWYLMDFSDY IASFVVAIII AHFGSKGNRT RWIAASCIL MGLESMLFAF PFFTYEIIIP GRQSIELCME ENEKRNIICG NSVPNRSKCI YFHIGGQCIH GIAGMPIYIL G ITFIFDHI PTSSCGFYLA IGHSAYLIGY LLGMVGGLQN FQPPPKEKTV EIEPAKVYQL LQSGWWKTFL IIAAISFCVS FM MVCFPTS LPGAHKLRLA KRKEPPTIDR RLKDMKIQPH LKGFLHNIWH ILKNPLMLTQ AICKVSEYLT FNTSLYFLPH HLQ TQFLIT PGIASLLTGA FVLPGGIIGH FLGGLIVDRL EMTNKNKLKF TLVTTVVSVG LFLLIFFVEC QTTTFAGINE DYDG YGQLG NLTADCNEYC DCTTSLYTSI CGRDEKEYFS PCFAGCKATK VSQTEKTYYN CSCIKEGLAA SDDEGQFIDA IAGTC DSDC LKLPLFFAFY FSATVFSNMC SIPVISIILQ SVPANFTSLS LGVTYAIVKF VASVPAPLLF RLSSAIACIY WDNNRC GGK ERCWIYNKNI LVYEFMGIWM SSQLIIVLLN IYAIQIHDVV VHGEITE

UniProtKB: Solute carrier organic anion transporter family member 6C1

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Macromolecule #13: Cation channel sperm-associated auxiliary subunit TMEM262

MacromoleculeName: Cation channel sperm-associated auxiliary subunit TMEM262
type: protein_or_peptide / ID: 13 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 13.350882 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString:
MRWRDRIAVL CFPPGLMLTV AALILFFIHM GVFASDVHNF CVIHNYDHMS FRYTVVLIFS QVISIGWAAM GSLYAEMTGD KFLRCFALT ILILNGAMFF NRLCLEFLAI NYREER

UniProtKB: Cation channel sperm-associated auxiliary subunit TMEM262

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Macromolecule #14: Unknown protein

MacromoleculeName: Unknown protein / type: protein_or_peptide / ID: 14 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 2.400951 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)

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Macromolecule #20: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 20 / Number of copies: 27 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

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Macromolecule #21: CHOLESTEROL

MacromoleculeName: CHOLESTEROL / type: ligand / ID: 21 / Number of copies: 3 / Formula: CLR
Molecular weightTheoretical: 386.654 Da
Chemical component information

ChemComp-CLR:
CHOLESTEROL

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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

BufferpH: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: INSILICO MODEL
Final reconstructionResolution.type: BY AUTHOR / Resolution: 6.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC / Number images used: 69708
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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