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- EMDB-32019: Cryo-EM structure of SAM-Tom40 intermediate complex -

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Basic information

Entry
Database: EMDB / ID: EMD-32019
TitleCryo-EM structure of SAM-Tom40 intermediate complex
Map data
Sample
  • Complex: SAM-Tom40
    • Protein or peptide: Sorting assembly machinery 50 kDa subunit
    • Protein or peptide: Sorting assembly machinery 35 kDa subunit
    • Protein or peptide: Sorting assembly machinery 37 kDa subunit
    • Protein or peptide: Mitochondrial import receptor subunit TOM40
Function / homology
Function and homology information


mitochondrial outer membrane translocase complex assembly / SAM complex / mitochondrial outer membrane translocase complex / protein import into mitochondrial matrix / phospholipid transport / protein insertion into mitochondrial outer membrane / porin activity / pore complex / protein transmembrane transporter activity / monoatomic ion transport ...mitochondrial outer membrane translocase complex assembly / SAM complex / mitochondrial outer membrane translocase complex / protein import into mitochondrial matrix / phospholipid transport / protein insertion into mitochondrial outer membrane / porin activity / pore complex / protein transmembrane transporter activity / monoatomic ion transport / mitochondrion organization / mitochondrial intermembrane space / protein-containing complex assembly / mitochondrial outer membrane / mitochondrion / cytosol / cytoplasm
Similarity search - Function
Sorting assembly machinery 35kDa subunit / Tom37, C-terminal domain / SAM35, subunit of SAM coomplex / Tom37 C-terminal domain / Mitochondrial import receptor subunit Tom40, fungi / Mitochondrial outer membrane transport complex Sam37/metaxin, N-terminal domain / Outer mitochondrial membrane transport complex protein / Tom40 / Eukaryotic porin/Tom40 / Eukaryotic porin ...Sorting assembly machinery 35kDa subunit / Tom37, C-terminal domain / SAM35, subunit of SAM coomplex / Tom37 C-terminal domain / Mitochondrial import receptor subunit Tom40, fungi / Mitochondrial outer membrane transport complex Sam37/metaxin, N-terminal domain / Outer mitochondrial membrane transport complex protein / Tom40 / Eukaryotic porin/Tom40 / Eukaryotic porin / Surface antigen D15-like / Bacterial surface antigen (D15) / Omp85 superfamily domain / Porin domain superfamily
Similarity search - Domain/homology
Sorting assembly machinery 35 kDa subunit / Mitochondrial import receptor subunit TOM40 / Sorting assembly machinery 37 kDa subunit / Sorting assembly machinery 50 kDa subunit
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) / Saccharomyces cerevisiae S288C (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsTakeda H / Tsutsumi A / Kikkawa M / Endo T
Funding support Japan, 3 items
OrganizationGrant numberCountry
Japan Society for the Promotion of Science (JSPS) Japan
Japan Science and Technology Japan
Japan Agency for Medical Research and Development (AMED) Japan
CitationJournal: To Be Published
Title: Cryo-EM structure of SAM-Tom40 intermediate complex
Authors: Takeda H / Tsutsumi A / Kikkawa M / Endo T
History
DepositionOct 1, 2021-
Header (metadata) releaseJan 4, 2023-
Map releaseJan 4, 2023-
UpdateJan 4, 2023-
Current statusJan 4, 2023Processing site: PDBj / Status: Released

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Structure visualization

Supplemental images

emd_32019.png

Downloads & links

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Map

FileDownload / File: emd_32019.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.245 Å
Density
Contour LevelBy AUTHOR: 0.06
Minimum - Maximum-0.20191672 - 0.42213887
Average (Standard dev.)-0.0002914935 (±0.0096141)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions200200200
Spacing200200200
CellA=B=C: 249.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : SAM-Tom40

EntireName: SAM-Tom40
Components
  • Complex: SAM-Tom40
    • Protein or peptide: Sorting assembly machinery 50 kDa subunit
    • Protein or peptide: Sorting assembly machinery 35 kDa subunit
    • Protein or peptide: Sorting assembly machinery 37 kDa subunit
    • Protein or peptide: Mitochondrial import receptor subunit TOM40

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Supramolecule #1: SAM-Tom40

SupramoleculeName: SAM-Tom40 / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast)

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Macromolecule #1: Sorting assembly machinery 50 kDa subunit

MacromoleculeName: Sorting assembly machinery 50 kDa subunit / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 54.47384 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MTSSSGVDNE ISLDSPMPIF NESSTLKPIR VAGVVTTGTD HIDPSVLQAY LDDTIMKSIT LGQLVKNADV LNKRLCQHHI ALNAKQSFH FQGNTYISDE KETHDVVPLM EVVSQLDILP PKTFTAKTGT NFGNDNDAEA YLQFEKLIDK KYLKLPTRVN L EILRGTKI ...String:
MTSSSGVDNE ISLDSPMPIF NESSTLKPIR VAGVVTTGTD HIDPSVLQAY LDDTIMKSIT LGQLVKNADV LNKRLCQHHI ALNAKQSFH FQGNTYISDE KETHDVVPLM EVVSQLDILP PKTFTAKTGT NFGNDNDAEA YLQFEKLIDK KYLKLPTRVN L EILRGTKI HSSFLFNSYS SLSPQSILNL KVFSQFYNWN TNKGLDIGQR GARLSLRYEP LFLHKLLHNP HSNESPTLFH EW FLETCWR STKICSQGTS APYMYSGTML SQAGDQLRTI LGHTFVLDKR DHIMCPTKGS MLKWSNELSP GKHLKTQLEL NSV KSWMND DFITFSTTIK TGYLKNLSSQ QSLPVHICDK FQSGGPSDIR GFQTFGLGPR DLYDAVGGDA FVSYGLSVFS RLPW KKVEK SNFRLHWFFN GGKLVNHDNT SLGNCIGQLS KEHSTSTGIG LVLRHPMARF ELNFTLPITA HENDLIRKGF QFGLG LAFL

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Macromolecule #2: Sorting assembly machinery 35 kDa subunit

MacromoleculeName: Sorting assembly machinery 35 kDa subunit / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 37.44607 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MVSSFSVPMP VKRIFDTFPL QTYAAQTDKD EAVALEIQRR SYTFTERGGG SSELTVEGTY KLGVYNVFLE ANTGAALATD PWCLFVQLA LCQKNGLVLP THSQEQTPSH TCNHEMLVLS RLSNPDEALP ILVEGYKKRI IRSTVAISEI MRSRILDDAE Q LMYYTLLD ...String:
MVSSFSVPMP VKRIFDTFPL QTYAAQTDKD EAVALEIQRR SYTFTERGGG SSELTVEGTY KLGVYNVFLE ANTGAALATD PWCLFVQLA LCQKNGLVLP THSQEQTPSH TCNHEMLVLS RLSNPDEALP ILVEGYKKRI IRSTVAISEI MRSRILDDAE Q LMYYTLLD TVLYDCWITQ IIFCASDAQF MELYSCQKLS GSIVTPLDVE NSLLQKLSAK SLKISLTKRN KFQFRHREIV KS MQGVYHN HHNSVNQEQV LNVLFENSKQ VLLGLKDMLK SDGQPTYLHL KIASYILCIT NVKEPIKLKT FVENECKELV QFA QDTLKN FVQ

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Macromolecule #3: Sorting assembly machinery 37 kDa subunit

MacromoleculeName: Sorting assembly machinery 37 kDa subunit / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 37.537797 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MVKGSVHLWG KDGKASLISV DSIALVWFIK LCTSEEAKSM VAGLQIVFSN NTDLSSDGKL PVLILDNGTK VSGYVNIVQF LHKNICTSK YEKGTDYEED LAIVRKKDRL LEYSLLNYVD VEISRLTDYQ LFLNTKNYNE YTKKLFSKLL YFPMWYNTPL Q LRSQAREN ...String:
MVKGSVHLWG KDGKASLISV DSIALVWFIK LCTSEEAKSM VAGLQIVFSN NTDLSSDGKL PVLILDNGTK VSGYVNIVQF LHKNICTSK YEKGTDYEED LAIVRKKDRL LEYSLLNYVD VEISRLTDYQ LFLNTKNYNE YTKKLFSKLL YFPMWYNTPL Q LRSQAREN CEEIIGSLTL EDDEEFVESK AMESASQLAQ SKTFKIAHKN KIKGKQELQQ VKYNLQFDNR LQSCVSNWLA AR KKLDDSV ILSSDLLFLA NLYVQLGLPD GNRIRSKLEQ TFGSELLNSM SNKIDDFVHR PSNNLEQRDP QFREQGNVVM SLY NLACKY I

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Macromolecule #4: Mitochondrial import receptor subunit TOM40

MacromoleculeName: Mitochondrial import receptor subunit TOM40 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 42.071141 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MSAPTPLAEA SQIPTIPALS PLTAKQSKGN FFSSNPISSF VVDTYKQLHS HRQSLELVNP GTVENLNKEV SRDVFLSQYF FTGLRADLN KAFSMNPAFQ TSHTFSIGSQ ALPKYAFSAL FANDNLFAQG NIDNDLSVSG RLNYGWDKKN ISKVNLQISD G QPTMCQLE ...String:
MSAPTPLAEA SQIPTIPALS PLTAKQSKGN FFSSNPISSF VVDTYKQLHS HRQSLELVNP GTVENLNKEV SRDVFLSQYF FTGLRADLN KAFSMNPAFQ TSHTFSIGSQ ALPKYAFSAL FANDNLFAQG NIDNDLSVSG RLNYGWDKKN ISKVNLQISD G QPTMCQLE QDYQASDFSV NVKTLNPSFS EKGEFTGVAV ASFLQSVTPQ LALGLETLYS RTDGSAPGDA GVSYLTRYVS KK QDWIFSG QLQANGALIA SLWRKVAQNV EAGIETTLQA GMVPITDPLM GTPIGIQPTV EGSTTIGAKY EYRQSVYRGT LDS NGKVAC FLERKVLPTL SVLFCGEIDH FKNDTKIGCG LQFETAGNQE LLMLQQGLDA DGNPLQALPQ L

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation statecell

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Sample preparation

BufferpH: 7.5
VitrificationCryogen name: ETHANE

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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: 1.8 µm / Nominal defocus min: 0.8 µm
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 48.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 23542
FSC plot (resolution estimation)

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