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- EMDB-31076: Structure of the CatSpermasome -

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Basic information

Entry
Database: EMDB / ID: EMD-31076
TitleStructure of the CatSpermasome
Map dataMap1, overall map, 2.9A
Sample
  • Complex: mouse CatSpermasome
    • Protein or peptide: x 14 types
  • Ligand: x 3 types
Function / homology
Function and homology information


CatSper complex / Sperm Motility And Taxes / sodium-independent organic anion transmembrane transporter activity / sodium-independent organic anion transport / regulation of cilium beat frequency involved in ciliary motility / sperm principal piece / fusion of sperm to egg plasma membrane involved in single fertilization / calcium ion sensor activity / flagellated sperm motility / male meiotic nuclear division ...CatSper complex / Sperm Motility And Taxes / sodium-independent organic anion transmembrane transporter activity / sodium-independent organic anion transport / regulation of cilium beat frequency involved in ciliary motility / sperm principal piece / fusion of sperm to egg plasma membrane involved in single fertilization / calcium ion sensor activity / flagellated sperm motility / male meiotic nuclear division / multicellular organism development / organic anion transport / organic anion transmembrane transporter activity / voltage-gated monoatomic ion channel activity / calcium-activated cation channel activity / regulation of monoatomic ion transmembrane transport / fertilization / motile cilium / sperm capacitation / monoatomic ion channel complex / sodium ion transport / plasma membrane => GO:0005886 / regulation of calcium ion transport / voltage-gated calcium channel activity / sperm flagellum / bioluminescence / acrosomal vesicle / generation of precursor metabolites and energy / calcium channel activity / cilium / calcium ion transport / spermatogenesis / cell differentiation / calmodulin binding / 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 zeta / Cation channel sperm-associated auxiliary subunit TMEM262 / EF-hand calcium-binding domain-containing protein 9 ...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 zeta / Cation channel sperm-associated auxiliary subunit TMEM262 / EF-hand calcium-binding domain-containing protein 9 / Cation channel sperm-associated protein subunit delta / Cation channel sperm-associated protein subunit gamma / Cation channel sperm-associated protein subunit beta protein family / Cation channel sperm-associated auxiliary subunit TMEM249 / Organic anion transporter polypeptide / Organic Anion Transporter Polypeptide (OATP) family / Kazal-type serine protease inhibitor domain / Kazal domain superfamily / Kazal domain / Kazal domain profile. / MFS transporter superfamily / Voltage-dependent channel domain superfamily / Green fluorescent protein, GFP / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / EF-hand calcium-binding domain profile. / Ion transport domain / Ion transport protein / EF-hand domain / EF-hand domain pair
Similarity search - Domain/homology
Transmembrane protein 249 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated protein subunit beta / Enhanced green fluorescent protein / Cation channel sperm-associated protein subunit gamma 2 / Transmembrane protein 262 / Cation channel sperm-associated protein subunit delta / Cation channel sperm-associated protein subunit epsilon / Kazal-like domain-containing protein / Cation channel sperm-associated protein 3 ...Transmembrane protein 249 / Cation channel sperm-associated protein 2 / Cation channel sperm-associated protein subunit beta / Enhanced green fluorescent protein / Cation channel sperm-associated protein subunit gamma 2 / Transmembrane protein 262 / Cation channel sperm-associated protein subunit delta / Cation channel sperm-associated protein subunit epsilon / Kazal-like domain-containing protein / Cation channel sperm-associated protein 3 / Cation channel sperm-associated protein 4 / Cation channel sperm-associated protein 1 / Cation channel sperm-associated protein subunit zeta / EF-hand calcium-binding domain-containing protein 9
Similarity search - Component
Biological speciesMus musculus (house mouse) / Mouse (mice)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.9 Å
AuthorsWu JP / Ke M
CitationJournal: Nature / Year: 2021
Title: Structure of a mammalian sperm cation channel complex.
Authors: Shiyi Lin / Meng Ke / Yuqi Zhang / Zhen Yan / Jianping Wu /
Abstract: The cation channel of sperm (CatSper) is essential for sperm motility and fertility. CatSper comprises the pore-forming proteins CATSPER1-4 and multiple auxiliary subunits, including CATSPERβ, γ, ...The cation channel of sperm (CatSper) is essential for sperm motility and fertility. CatSper comprises the pore-forming proteins CATSPER1-4 and multiple auxiliary subunits, including CATSPERβ, γ, δ, ε, ζ, and EFCAB9. Here we report the cryo-electron microscopy (cryo-EM) structure of the CatSper complex isolated from mouse sperm. In the extracellular view, CATSPER1-4 conform to the conventional domain-swapped voltage-gated ion channel fold, following a counterclockwise arrangement. The auxiliary subunits CATSPERβ, γ, δ and ε-each of which contains a single transmembrane segment and a large extracellular domain-constitute a pavilion-like structure that stabilizes the entire complex through interactions with CATSPER4, 1, 3 and 2, respectively. Our EM map reveals several previously uncharacterized components, exemplified by the organic anion transporter SLCO6C1. We name this channel-transporter ultracomplex the CatSpermasome. The assembly and organization details of the CatSpermasome presented here lay the foundation for the development of CatSpermasome-related treatments for male infertility and non-hormonal contraceptives.
History
DepositionMar 18, 2021-
Header (metadata) releaseJul 28, 2021-
Map releaseJul 28, 2021-
UpdateAug 11, 2021-
Current statusAug 11, 2021Processing site: PDBj / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.4
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by height
  • Surface level: 0.4
  • Imaged by UCSF Chimera
  • Download
  • Surface view with fitted model
  • Atomic models: PDB-7eeb
  • Surface level: 0.4
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_31076.png

Downloads & links

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Map

FileDownload / File: emd_31076.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMap1, overall map, 2.9A
Voxel sizeX=Y=Z: 1.0773 Å
Density
Contour LevelBy AUTHOR: 0.4 / Movie #1: 0.4
Minimum - Maximum-2.3705251 - 4.092949
Average (Standard dev.)0.0025662975 (±0.08306798)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 430.91998 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.07731.07731.0773
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z430.920430.920430.920
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ400400400
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS400400400
D min/max/mean-2.3714.0930.003

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Supplemental data

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Additional map: Map2, improved map quality near CatSper eta

Fileemd_31076_additional_1.map
AnnotationMap2, improved map quality near CatSper eta
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: Map3, improved map quality near SLCO6C1

Fileemd_31076_additional_2.map
AnnotationMap3, improved map quality near SLCO6C1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: Map4, improved map quality near CatSper delta-TM

Fileemd_31076_additional_3.map
AnnotationMap4, improved map quality near CatSper delta-TM
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: Map5, improved map quality in cytosolic region

Fileemd_31076_additional_4.map
AnnotationMap5, improved map quality in cytosolic region
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: composite map for interpretation

Fileemd_31076_additional_5.map
Annotationcomposite map for interpretation
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : mouse CatSpermasome

EntireName: mouse CatSpermasome
Components
  • Complex: mouse CatSpermasome
    • Protein or peptide: Enhanced green fluorescent protein,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 protein subunit beta
    • Protein or peptide: Cation channel sperm-associated protein subunit gamma 2
    • Protein or peptide: Cation channel sperm-associated protein subunit delta
    • Protein or peptide: Cation channel sperm-associated protein subunit epsilon
    • Protein or peptide: Kazal-like domain-containing protein
    • Protein or peptide: Transmembrane protein 249Transmembrane protein
    • Protein or peptide: Transmembrane protein 262Transmembrane protein
    • Protein or peptide: Unknown
    • Protein or peptide: EF-hand calcium-binding domain-containing protein 9
    • Protein or peptide: Cation channel sperm-associated protein subunit zeta
  • Ligand: SODIUM IONSodium
  • Ligand: (3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]spirost-5-en
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose

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Supramolecule #1: mouse CatSpermasome

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

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Macromolecule #1: Enhanced green fluorescent protein,Cation channel sperm-associate...

MacromoleculeName: Enhanced green fluorescent protein,Cation channel sperm-associated protein 1
type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mus musculus (house mouse)
Molecular weightTheoretical: 109.442758 KDa
Recombinant expressionOrganism: Mus musculus (house mouse)
SequenceString: DYKDHDGDYK DHDIDYKDDD DKMVSKGEEL FTGVVPILVE LDGDVNGHKF SVSGEGEGDA TYGKLTLKFI CTTGKLPVPW PTLVTTLTY GVQCFSRYPD HMKQHDFFKS AMPEGYVQER TIFFKDDGNY KTRAEVKFEG DTLVNRIELK GIDFKEDGNI L GHKLEYNY ...String:
DYKDHDGDYK DHDIDYKDDD DKMVSKGEEL FTGVVPILVE LDGDVNGHKF SVSGEGEGDA TYGKLTLKFI CTTGKLPVPW PTLVTTLTY GVQCFSRYPD HMKQHDFFKS AMPEGYVQER TIFFKDDGNY KTRAEVKFEG DTLVNRIELK GIDFKEDGNI L GHKLEYNY NSHNVYIMAD KQKNGIKVNF KIRHNIEDGS VQLADHYQQN TPIGDGPVLL PDNHYLSTQS ALSKDPNEKR DH MVLLEFV TAAGITLGMD ELYKENLYFQ GSMDQSSRRD ESYHETHPGS LDPSHQSHPH PHPHPTLHRP NQGGVYYDSP QHG MFQQPY QQHGGFHQQN ELQHLREFSD SHDNAFSHHS YQQDRAGVST LPNNISHAYG GSHPLAESQH SGGPQSGPRI DPNH HPHQD DPHRPSEPLS HPSSTGSHQG TTHQQYHERS HHLNPQQNRD HADTISYRSS TRFYRSHAPF SRQERPHLHA DHHHE GHHA HSHHGEHPHH KEQRHYHGDH MHHHIHHRSP SASQLSHKSH STLATSPSHV GSKSTASGAR YTFGARSQIF GKAQSR ESL RESASLSEGE DHVQKRKKAQ RAHKKAHTGN IFQLLWEKIS HLLLGLQQMI LSLTQSLGFE TFIFIVVCLN TVILVAQ TF TELEIRGEWY FMVLDSIFLS IYVLEAVLKL IALGLEYFYD PWNNLDFFIM VMAVLDFVLL QINSLSYSFY NHSLFRIL K VFKSMRALRA IRVLRRLSIL TSLHEVAGTL SGSLPSITAI LTLMFTCLFL FSVVLRALFQ DSDPKRFQNI FTTLFTLFT MLTLDDWSLI YIDNRAQGAW YIIPILMIYI VIQYFIFLNL VIAVLVDNFQ MALLKGLEKV KLEQAARVHE KLLDDSLTDL NKADANAQM TEEALKMQLI EGMFGNMTVK QRVLHFQFLQ LVAAVEQHQQ KFRSQAYVID ELVDMAFEAG DDDYGK

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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: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 68.651062 KDa
SequenceString: MAQEQGHFQL LRADAIRSKL IDTFSLIEHL QGLSQAVPRH TLREILDPSY QQKLMSGDQE QLVRFSIKPR RMGHITHSRR LLSRLRVRC SRMPPLSLWA GWVLDSSVFS KFIISLIFLN TFVLMVEIEL MESTNTALWP VKLALEVADW FILLSFIVEI L LMWLASFS ...String:
MAQEQGHFQL LRADAIRSKL IDTFSLIEHL QGLSQAVPRH TLREILDPSY QQKLMSGDQE QLVRFSIKPR RMGHITHSRR LLSRLRVRC SRMPPLSLWA GWVLDSSVFS KFIISLIFLN TFVLMVEIEL MESTNTALWP VKLALEVADW FILLSFIVEI L LMWLASFS LFWKDAWNVF DFFVTLLSLL PELVVLLGVP AHSVWLQLLR VCRVLRSLKL FARFRQIKVI LLALVRALKS MT FLLMLLL IFFYIFAVTG VYFFREYSRS TIEGLEYNMF FSDLLNSLVT VFILFTLDHW YAVLQDIWKV PESSRVFSSI YVI LWLLLG SIIFRNIIVA MMVTNFQNIR SELSEEMSHL EVQYKADMFK QQIIQRRQHS ESLRGTSLGK VSEDIIETSD ASDD DDDDD DDDDDDDDDD DDKSDATESD NEESDSENSE SENSESEKID PEKDYAKKSY PEKSHPEKSY PEKSHPEKSY PEKSH PKKS YDEQAEAEKV KEESKEKAYP VSHSISSHGS TAADTAFLEN LDWETLVHEN LPGLMDMDQD DRIVWPRDSL FRYFEL LEK LQYNLEERKK LQEFAVQALM SFEDK

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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: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 45.533113 KDa
SequenceString: MSQHFHHNPV RVKSGSLFAT ASEALQARLS KIKRKDKECQ AYFRKVIKST FFQIVMITTV TTNSFLLVLG TNYDIQFEFF RTFEVSELF FVSVYVCEFL MKVYVDPITY WKDGYNILDV IILIILTIPY LLRKIKGNHS AYLHFADGIQ SLRILKLISY S RGIRTLII ...String:
MSQHFHHNPV RVKSGSLFAT ASEALQARLS KIKRKDKECQ AYFRKVIKST FFQIVMITTV TTNSFLLVLG TNYDIQFEFF RTFEVSELF FVSVYVCEFL MKVYVDPITY WKDGYNILDV IILIILTIPY LLRKIKGNHS AYLHFADGIQ SLRILKLISY S RGIRTLII AVGETVYTVA SVLTLLFLLM FVFAILGFCL FGVTDRGDLE NWGNLASAFF TLFSLATVDG WTDLQEELDK RK FTVSRAF TILFILLASF IFLNMFVGVM IMHTEDSMKK FERDLTLERN LAIMEEKQII LKRQQEEVNR LMNTQKSGSM NFI DMVEGF KKTLRHTDPM VLDDFSTSLS FIDIYLVTLD NQDVIVSKLQ ELYCEIVNVL SLMLEDMPKE SSSSLSGLS

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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: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 51.179273 KDa
SequenceString: MSEKHKWWQQ VENIDITHLG PKRKAYELLG RHEEQVLINR RDVMEKKDAW DVQEFITQMY IKQLLRHPAF QLLLAFLLLS NAITIALRT NSYLGQKHYE LFSTIDDIVL TILICEVLLG WLNGFWIFWK DGWNILNFAI VFILFMGFFI KQLDMVAITY P LRVLRLVH ...String:
MSEKHKWWQQ VENIDITHLG PKRKAYELLG RHEEQVLINR RDVMEKKDAW DVQEFITQMY IKQLLRHPAF QLLLAFLLLS NAITIALRT NSYLGQKHYE LFSTIDDIVL TILICEVLLG WLNGFWIFWK DGWNILNFAI VFILFMGFFI KQLDMVAITY P LRVLRLVH VCMAVEPLAR IIKVILQSMP DLANVMALIL FFMLVFSVFG VTLFGAFVPK HFQNMGVALY TLFICITQDG WL DIYTDFQ MDEREYAMEV GGAIYFAVFI TLGAFIGLNL FVVVVTTNLE QMMKTGEEEG HLNIKFTETE EDEDWTDELP LVH CTEARK DTSTVPKEPL VGGPLSNLTE KTCDNFCLVL EAIQENLMEY KEIREELNMI VEEVSSIRFN QEQQNVILHK YTSK SATFL SEPPEGANKQ DLITALVSRE KVSDSNINMV NKHKFSH

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

MacromoleculeName: Cation channel sperm-associated protein subunit beta / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 126.250031 KDa
SequenceString: MESPLIYVML VLLNVFVFSS GVIHNKGKER TYFSCSGEGI LTGLHTIKLF LTMDNLKVRC FFRNENQSPS KEILGLFTSG GLAPNMIIT NSTFYGGYYF KLTPFSNRLE WLIDIPRQNI TVNTDIAAVE QWMIKITMHE GLNIYDTEGT LLDLVREPIL Q WNLGRVLT ...String:
MESPLIYVML VLLNVFVFSS GVIHNKGKER TYFSCSGEGI LTGLHTIKLF LTMDNLKVRC FFRNENQSPS KEILGLFTSG GLAPNMIIT NSTFYGGYYF KLTPFSNRLE WLIDIPRQNI TVNTDIAAVE QWMIKITMHE GLNIYDTEGT LLDLVREPIL Q WNLGRVLT EMEVRDLYPE VNDIKVTKSP CANDVALIGF MMKPSSNGVF IGKTISGFWT YKECIWHDLT EIIYAELKDE HQ GLTVIDL VLTNHFLVIL TSLGLYVSSD LRYPTTSQIK LSRAEFCGFE RVDYIRGNLW YNEKCFANRE SFEVDYVTIT FNR NRTLSE SSSCFFSKEP FLHWLPCVFS TIKNEKSIPR VITFLIDQET DSGIYLFNVQ DTKETYVTVA MLKDGKPSPR PKFP SFHFP STFTLPLGMI FHPRSHFLYV YGSQIWVSMD GGNTFEMLCN LFSHHVTKTS NSFYTSDIVF IVEDGRILTT KAGLT TYSE LGILKDAIFT LYYDQLGYIH KLTPENFDAG SKLLGHGNSG SIFGKRPDLG FEAILVPQYI STNEMYFFAH VPLTMP TNI QWKKRFKTIH LGKTIEFSKT GLANIKNVYM HKTEPVGFQT SIHTEIIVPF GIENSKDSPC LLSDLEITYS GKLYYTI KL LSKNPLHELK STDVEKSVLI PGYSSFLIMN ITDKWTASAL ATMPQAIKSN LKFLTGSWFL YNFGTAGGRK WSISTRQC N YWIQQDSLDF MSLNLVKYID VGNTIDFQFK IIPKAMSTFP IPPVSMVVGN PGLVEVKTQG VFDLNENYYL DIHVSGRFF QKGSTSIALV LWEGSSKCYA ITLLPTIKSS CSYLRTMHHT PGRHIPPEDW ISGVHKDSQG FNMIKTLPIN YRPPSHMGIS IPLTDNFYH ADPSKPIPRN QFHKSKETGK YKQCANVTSR AMCNCSEHQK FSHAVAFSDC KEKVHRFKFP VTQYPVVLEI F NERDKISA EPPYLVTMTE VNMRKNWQLK HNEPENVKKM KHYLEPLLKT PVYNPLGLNL TIQGSELFHF KVSVVPGVSF CE LSEEFQI YVDEVPLPFP GHALIAVATS VVLGVLIFIA FVFQLRNIHP LKALKKSIRG NPGLTSSTTV SS

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

MacromoleculeName: Cation channel sperm-associated protein subunit gamma 2
type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 131.550594 KDa
SequenceString: MVSRPAMSPV SPVWPRKPNL WAFWVLRLVL LLSLKSWAED ALQHCTWLLV LNKFEKVGLH LSKDRFQDHE PIDTVAKVFQ KLTDSPIDP SENYLSFPYY LQINFSCPGQ NIEELARKGH LMGMKPMVQI NYMYSVNFYR WEMENVQILM EAAPMRSTGY C PAEAMCVL ...String:
MVSRPAMSPV SPVWPRKPNL WAFWVLRLVL LLSLKSWAED ALQHCTWLLV LNKFEKVGLH LSKDRFQDHE PIDTVAKVFQ KLTDSPIDP SENYLSFPYY LQINFSCPGQ NIEELARKGH LMGMKPMVQI NYMYSVNFYR WEMENVQILM EAAPMRSTGY C PAEAMCVL NWYTPMPFKN GSVVSSVDIY TNGIGPFVSK KRFYVNMNGF LKRDASGKSL FAIGYESLVL KSSHFRLSKS RP LWYTVNH APVFILGGFY DEKSILFSDS NFQDYVLLEL SIDSCWVGSF YCPILGFSAT IHDAIATEST LFIRQNQLVY YFT GTYITL FDKSHGSSRW VRVLPSECIK RLCPVYASGN GSEYVLALTT GKNEGYIHIG TITDGLVSFE MVPDGWSVCE KLPG KNCSI DWATYIADER NLLLLVKIDS GQFYLVNFNT EFKTLNILYK IPEFIPEAKE LDFLVLLDTV TYTNTPMTPK GLFFN TLNN MLYIWGNFIL QSYNREEFIF LADFPKESTI KYMVNSFKGQ MAVVTENEEI WYFLEGGYDV YQVVPSQGWE TYHNLQ KMQ KSSFHSEDES LVSLFFEDGK LFQLVYLFDV GKERLVKRLL PVGTLMEYNL PKPFTVVNQG NYQAISFTHT CPFKEIH LI DVPKKHHASR TESYVALPPL VSESLGFHNN NTLAVYQGLV YYLLWLHSKY DKPYADPVHD PTWRWWQHKT KDKDYFFY L FSNRLAAEGI YINMNAYQKL YNMSGDYGIP DLFFLDKGNW FTITVVLLSH QDTFTSSDSQ GPTINVDKKL AIAVTIADP ECLSVTVTQD VLLNRNAVIN KIKVIDKKRC SEQGMIGRNI KKTSMMLKVL GAPGNCIQRT YLGGIIQGFK VVPIFIGCPP GKRLAFDVS YTIMHSEEIN KHYFDCVIKD AEMPCFLFRD LFQPFFLVQD LVTGDSGSFL GSYVLKVVGG GRTLNTIRDY T EEEIFRYN SPLDTTNSLI WKTKVERTTE DKKFYIMSHE SPGVEWLCLE NSPCYDIIPQ SIYPPEFFFK LLVSNRGVDN ST YCDYKLT FIVHIHGLPL SSKRTSFIVM VSTSFFIALV VFYILFCLVW PHIVKAWVSL RWRINNIMAS ESYYTYASST AGF SLQSHS FEGPSRAGSK EDNVQAKTA

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

MacromoleculeName: Cation channel sperm-associated protein subunit delta / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 91.186453 KDa
SequenceString: MLVLMLAAAV ATMVRAHTLC RVHTVRTGKV FKSNIQLQGD PLFYAFPNTF VLKNVCKADI SVYLGQKVFL TIDNFESSLL PLTVPKSLA VGVPSITSAH FVSGSLVLFV ISGKGYSYDY YENTWRKLEG ISEPVSHISG DVCCFKGSFC LELSNNLFAY L RGGQIPGT ...String:
MLVLMLAAAV ATMVRAHTLC RVHTVRTGKV FKSNIQLQGD PLFYAFPNTF VLKNVCKADI SVYLGQKVFL TIDNFESSLL PLTVPKSLA VGVPSITSAH FVSGSLVLFV ISGKGYSYDY YENTWRKLEG ISEPVSHISG DVCCFKGSFC LELSNNLFAY L RGGQIPGT NIYFSDNGGF SFQLMNTDKL SHLTGTLGGI FHLHSMSQVG VLMVENNLGT FHYMEYPLNH SMGIAFSYKN LL EVIMKPY QRGFMVLWNQ KSILVSSNSG QIVEHVRLID QKIFTDLDVE HANINIYSVA SNAYELAFLV AEDHLYYGSQ SYM GTYVIK LPHQPLWSTH TSIYFEDIGI LQVLTPVADP HFAAYDFDKC TVNVQSSLMD EKLALQPCNV ELLESTMINT MFTI DMNSK LKLSALMIPR KGENPTPLVM VSNPHALGFK ANLNEFGNTF DGNSKYKLDI ELKQQHHWGN SDFNFTASIK RHAIS SVTV DIADKTLSCV DLKPLSTLIS VGCDMTKKIV VQNKISACTM GILNPVQLQK NYTYTIEKEA YDPINHNGEA QDDLIV FYE YKDLGCPRLV YYDKPWKPVV ELWKNGIVEE IMNAEYVISE INGLVTYSYS LTAATANCRS QPQNWSTFES DIENEEP FL WNRENYVSCH EDNKDNPLLW PNVEYQVLGG QTNNKIIFGQ RNGIYTFHLS VVDPYYSYCN LNTIFSVYVH GALPVTKF Q PLLTILLMVT TTLLTAWLAY AIPKQLRSEK GQRLLGFCYQ ILQLCLGVCF CTWLRGKLRQ WLRPRRVKDQ NRGKVRVAQ KHPET

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

MacromoleculeName: Cation channel sperm-associated protein subunit epsilon
type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 113.913453 KDa
SequenceString: MPSAGQRKPG SLLALQALQK WLLRGGVGAM LARQVVAALL LWLSCCVSAL WRYYINSQDY SIFSTRSSIK LEYEGNSFVS WKIPESCKV ENTTSPKTTL HCKRAGIHTI KPIAGNQEVE RHLTVDNSYI CYLWYFTVVD VYYNLSQIVT IWVYDPESAS T EELIWTAK ...String:
MPSAGQRKPG SLLALQALQK WLLRGGVGAM LARQVVAALL LWLSCCVSAL WRYYINSQDY SIFSTRSSIK LEYEGNSFVS WKIPESCKV ENTTSPKTTL HCKRAGIHTI KPIAGNQEVE RHLTVDNSYI CYLWYFTVVD VYYNLSQIVT IWVYDPESAS T EELIWTAK KPSLSSRVLT KQMNTLGQRP FIFTVEKRLT YHPGPLTSEG TWVIHLPMSS DDIAKVIRGN KVAFQDCFIA NL YFMLTYP MTIISEPPGY EPLTVPPGSP LMLSWDTCIS TFALLATDQE TFQTNDSFQT WTRVRAPPGI LSDAQRHSLR DVI IFDQGT LFLVDGTVYL RTEDEFTKLD ESRGISETGI LGFSKRRWCQ IRYLYKLASK KSILIAWSKT TVYAGYATFR FVTL TDTAK LKDFLKLPQT DTLEVMSVEY LWHPLEAAVL LSHCSVCTTN TRNIRIVIYS AIFQTWTLQD FELQLPKEAI LEFRF LYSA MPDIIMWDQH HVYYSYKNFT VVGTISTPSG ETNLSSLSQG SKIHQVLTDR IGNVVVKMEN NVMFYIKADI TEAVIL HTW VNTTAKTVVL FDKSFEVCIL YYNENLDEKY QLQTQPYPLI LELQSINKDL GDWCPYLAFQ HNIHSQFYHM DKGESLT IW SQIVYPENRG LYIVVEHYGS SVMTWTQNLE YEIASGFCTK TMITRFFQTT NYELVDNYYQ LQKENTGLML LQFRPSEF S RTCLTAKPVF EIDVGCDSSK YIMVRGFNKS RCQRRDFSYV IDKELLRESL SDNLKVRYDV AKYGCPLTLE LGQMFQPIV ELYDENGFIK IVDANFILWE IHGRNDYTFN STMEQNGCIN EAQTWDSMIE ENPDIPLDDV WGPQNYRPCF SYAIGKPGDL GQPYEILNY SNKNHIKWPM TYAGMYVYRL KILDPNYSFC NLTTIFAIES LGMIPRSSVY LVAALIFVLM LTFISILVLS Y FWYLKIYR QFIIEPLHKR PAKQKKN

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Macromolecule #9: Kazal-like domain-containing protein

MacromoleculeName: Kazal-like domain-containing protein / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 79.055258 KDa
SequenceString: MAHVRNKKSD DKKAMVVAKE DTNKSESEGV TKLQTYLKTI PIAKKKFAKL PKRKKSPTSA ELLLIDPRYS ASKEGPLGLG PIVLPFVQR FNNIDGFMTL YVAAVLIHGA LFAVVDMTLN IYQVQFSLTR TEWYLMDFSD YIASFVVAII IAHFGSKGNR T RWIAASCI ...String:
MAHVRNKKSD DKKAMVVAKE DTNKSESEGV TKLQTYLKTI PIAKKKFAKL PKRKKSPTSA ELLLIDPRYS ASKEGPLGLG PIVLPFVQR FNNIDGFMTL YVAAVLIHGA LFAVVDMTLN IYQVQFSLTR TEWYLMDFSD YIASFVVAII IAHFGSKGNR T RWIAASCI LMGLESMLFA FPFFTYEIII PGRQSIELCM EENEKRNIIC GNSVPNRSKC IYFHIGGQCI HGIAGMPIYI LG ITFIFDH IPTSSCGFYL AIGHSAYLIG YLLGMVGGLQ NFQPPPKEKT VEIEPAKVYQ LLQSGWWKTF LIIAAISFCV SFM MVCFPT SLPGAHKLRL AKRKEPPTID RRLKDMKIQP HLKGFLHNIW HILKNPLMLT QAICKVSEYL TFNTSLYFLP HHLQ TQFLI TPGIASLLTG AFVLPGGIIG HFLGGLIVDR LEMTNKNKLK FTLVTTVVSV GLFLLIFFVE CQTTTFAGIN EDYDG YGQL GNLTADCNEY CDCTTSLYTS ICGRDEKEYF SPCFAGCKAT KVSQTEKTYY NCSCIKEGLA ASDDEGQFID AIAGTC DSD CLKLPLFFAF YFSATVFSNM CSIPVISIIL QSVPANFTSL SLGVTYAIVK FVASVPAPLL FRLSSAIACI YWDNNRC GG KERCWIYNKN ILVYEFMGIW MSSQLIIVLL NIYAIQIHDV VVHGEITESK TTVKDVKEQK ERKA

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Macromolecule #10: Transmembrane protein 249

MacromoleculeName: Transmembrane protein 249 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 19.918379 KDa
SequenceString:
MLFIICLVFI SCNVLREVKY QETWCFPAYG MVIGLWLMLS SIPQRRLVLN HTRGMYHFSI QGRTVCQGPM HLVYVRLALS SDAYGGRFF QLVLCGHKLE PLVLVQLSER YEQMEFLGRH LARKLNINYF DYLASSYRHV VRHWPLGASF SPGIVQRKTQ V YTKSSVND LDV

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Macromolecule #11: Transmembrane protein 262

MacromoleculeName: Transmembrane protein 262 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 13.489028 KDa
SequenceString:
MRWRDRIAVL CFPPGLMLTV AALILFFIHM GVFASDVHNF CVIHNYDHMS FRYTVVLIFS QVISIGWAAM GSLYAEMTGD KFLRCFALT ILILNGAMFF NRLCLEFLAI NYREERH

+
Macromolecule #12: Unknown

MacromoleculeName: Unknown / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
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 #13: EF-hand calcium-binding domain-containing protein 9

MacromoleculeName: EF-hand calcium-binding domain-containing protein 9 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 26.170482 KDa
SequenceString: MKLTPGCFLW YLYMDKIYCL LSLRNVKALM VYFHLLDVHH RNTLNDVLFF HFLQHVTNLN KSQIGMIFDL LDWTAVGEIG FDQFYVLIC ILLAHQDHLE DHFMYRHSRP VFELLDLDGE MNIGAANFQN YRFLFNIKKQ ELRDLFHDFD ITGDRLLNYK E FKLYTIFC ...String:
MKLTPGCFLW YLYMDKIYCL LSLRNVKALM VYFHLLDVHH RNTLNDVLFF HFLQHVTNLN KSQIGMIFDL LDWTAVGEIG FDQFYVLIC ILLAHQDHLE DHFMYRHSRP VFELLDLDGE MNIGAANFQN YRFLFNIKKQ ELRDLFHDFD ITGDRLLNYK E FKLYTIFC TDKSIDRKKR RKDREAARER EKEKGKDKEK YLHLKKIYSS MLSHRSIL

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Macromolecule #14: Cation channel sperm-associated protein subunit zeta

MacromoleculeName: Cation channel sperm-associated protein subunit zeta / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Mouse (mice)
Molecular weightTheoretical: 22.776598 KDa
SequenceString: MEESVKPVPK HANHRRSSVR SSLYGDVRDL WSTATMSTAN VSVSDVCEDF DEEGKSVRNR IRKYSQTISI RDSLNLEPEE IQQQARREL ELCHGRSLEH GEDHEESETS LASSTSESLI FSLWKPHRTY WTEQQNRLPL PLMELMETEV LDILKKALIT Y RSTIGRNH ...String:
MEESVKPVPK HANHRRSSVR SSLYGDVRDL WSTATMSTAN VSVSDVCEDF DEEGKSVRNR IRKYSQTISI RDSLNLEPEE IQQQARREL ELCHGRSLEH GEDHEESETS LASSTSESLI FSLWKPHRTY WTEQQNRLPL PLMELMETEV LDILKKALIT Y RSTIGRNH FMTKELQGYI EGIRKRRNKR LYFLDQ

+
Macromolecule #20: SODIUM ION

MacromoleculeName: SODIUM ION / type: ligand / ID: 20 / Number of copies: 2
Molecular weightTheoretical: 22.99 Da

+
Macromolecule #21: (3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]sp...

MacromoleculeName: (3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]spirost-5-en
type: ligand / ID: 21 / Number of copies: 3 / Formula: 9Z9
Molecular weightTheoretical: 544.805 Da
Chemical component information

ChemComp-9Z9:
(3beta,14beta,17beta,25R)-3-[4-methoxy-3-(methoxymethyl)butoxy]spirost-5-en / detergent*YM

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Macromolecule #22: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 22 / Number of copies: 9 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

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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
GridModel: Homemade / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 281 K / Instrument: FEI VITROBOT MARK IV / Details: blot for 8 s before plunging.

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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 defocus max: 2.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

-
Image processing

CTF correctionSoftware - Name: cryoSPARC (ver. v3.0)
Initial angle assignmentType: NOT APPLICABLE
Final angle assignmentType: NOT APPLICABLE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 560730

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