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- EMDB-28744: Cryo-EM consensus map of the S. cerevisiae Arf-like protein Arl1 ... -

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Basic information

Entry
Database: EMDB / ID: EMD-28744
TitleCryo-EM consensus map of the S. cerevisiae Arf-like protein Arl1 bound to its effector guanine nucleotide exchange factor Gea2
Map data
Sample
  • Complex: Gea2 homodimer in complex with two Arl1 monomers
    • Protein or peptide: ARF guanine-nucleotide exchange factor 2
    • Protein or peptide: ADP-ribosylation factor-like protein 1
Keywordssmall GTPase / guanine nucleotide exchange factor / membrane trafficking / lipid flippase / trans-Golgi network / protein transport / LIPID TRANSPORT
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.3 Å
AuthorsDuan HD / Li H
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Cancer Institute (NIH/NCI)CA231466 United States
CitationJournal: To Be Published
Title: Structural insights into the recruitment of ArfGEF Gea2 by Arl1 to the Drs2-localizing trans-Golgi network
Authors: Duan HD / Li H
History
DepositionNov 1, 2022-
Header (metadata) releaseNov 15, 2023-
Map releaseNov 15, 2023-
UpdateNov 15, 2023-
Current statusNov 15, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_28744.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.828 Å
Density
Contour LevelBy AUTHOR: 0.01
Minimum - Maximum-0.027672121 - 1.9048703
Average (Standard dev.)0.0006522556 (±0.016357344)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions512512512
Spacing512512512
CellA=B=C: 423.936 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_28744_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: #1

Fileemd_28744_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire : Gea2 homodimer in complex with two Arl1 monomers

EntireName: Gea2 homodimer in complex with two Arl1 monomers
Components
  • Complex: Gea2 homodimer in complex with two Arl1 monomers
    • Protein or peptide: ARF guanine-nucleotide exchange factor 2
    • Protein or peptide: ADP-ribosylation factor-like protein 1

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Supramolecule #1: Gea2 homodimer in complex with two Arl1 monomers

SupramoleculeName: Gea2 homodimer in complex with two Arl1 monomers / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Macromolecule #1: ARF guanine-nucleotide exchange factor 2

MacromoleculeName: ARF guanine-nucleotide exchange factor 2 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MHHHHHHSSG VDLGTENLYF QSNAMSDREF VTVDPVTIII KECINLSTAM RKYSKFTSQS GVAALLGGGS EIFSNQDDYL AHTFNNLNT NKHNDPFLSG FIQLRLMLNK LKNLDNIDSL TILQPFLLIV STSSISGYIT SLALDSLQKF FTLNIINESS Q NYIGAHRA ...String:
MHHHHHHSSG VDLGTENLYF QSNAMSDREF VTVDPVTIII KECINLSTAM RKYSKFTSQS GVAALLGGGS EIFSNQDDYL AHTFNNLNT NKHNDPFLSG FIQLRLMLNK LKNLDNIDSL TILQPFLLIV STSSISGYIT SLALDSLQKF FTLNIINESS Q NYIGAHRA TVNALTHCRF EGSQQLSDDS VLLKVVFLLR SIVDSPYGDL LSNSIIYDVL QTILSLACNN RRSEVLRNAA QS TMIAVTV KIFSKLKTIE PVNVNQIYIN DESYTNDVLK ADTIGTNVES KEEGSQEDPI GMKVNNEEAI SEDDGIEEEH IHS EKSTNG AEQLDIVQKT TRSNSRIQAY ADDNYGLPVV RQYLNLLLSL IAPENELKHS YSTRIFGLEL IQTALEISGD RLQL YPRLF TLISDPIFKS ILFIIQNTTK LSLLQATLQL FTTLVVILGN NLQLQIELTL TRIFSILLDD GTANNSSSEN KNKPS IIKE LLIEQISILW TRSPSFFTST FINFDCNLDR ADVSINFLKA LTKLALPESA LTTTESVPPI CLEGLVSLVD DMFDHM KDI DREEFGRQKN EMEILKKRDR KTEFIECTNA FNEKPKKGIP MLIEKGFIAS DSDKDIAEFL FNNNNRMNKK TIGLLLC HP DKVSLLNEYI RLFDFSGLRV DEAIRILLTK FRLPGESQQI ERIIEAFSSA YCENQDYDPS KISDNAEDDI STVQPDAD S VFILSYSIIM LNTDLHNPQV KEHMSFEDYS GNLKGCCNHK DFPFWYLDRI YCSIRDKEIV MPEEHHGNEK WFEDAWNNL ISSTTVITEI KKDTQSVMDK LTPLELLNFD RAIFKQVGPS IVSTLFNIYV VASDDHISTR MITSLDKCSY ISAFFDFKDL FNDILNSIA KGTTLINSSH DDELSTLAFE YGPMPLVQIK FEDTNTEIPV STDAVRFGRS FKGQLNTVVF FRIIRRNKDP K IFSKELWL NIVNIILTLY EDLILSPDIF PDLQKRLKLS NLPKPSPEIS INKSKESKGL LSTFASYLKG DEEPTEEEIK SS KKAMECI KSSNIAASVF GNESNITADL IKTLLDSAKT EKNADNSRYF EAELLFIIEL TIALFLFCKE EKELGKFILQ KVF QLSHTK GLTKRTVRRM LTYKILLISL CADQTEYLSK LINDELLKKG DIFTQKFFAT NQGKEFLKRL FSLTESEFYR GFLL GNENF WKFLRKVTAM KEQSESIFEY LNESIKTDSN ILTNENFMWV LGLLDEISSM GAVGNHWEIE YKKLTESGHK IDKEN PYKK SIELSLKSIQ LTSHLLEDNN DLRKNEIFAI IQALAHQCIN PCKQISEFAV VTLEQTLINK IEIPTNEMES VEELIE GGL LPLLNSSETQ EDQKILISSI LTIISNVYLH YLKLGKTSNE TFLKILSIFN KFVEDSDIEK KLQQLILDKK SIEKGNG SS SHGSAHEQTP ESNDVEIEAT APIDDNTDDD NKPKLSDVEK D

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Macromolecule #2: ADP-ribosylation factor-like protein 1

MacromoleculeName: ADP-ribosylation factor-like protein 1 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
ELRILILGLD GAGKTTILYR LQIGEVVTTK PTIGFNVETL SYKNLKLNVW DLGGLTSIRP YWRCYYADTA AVIFVVDSTD KDRMSTASK ELHLMLQEEE LQDAALLVFA NKQDQPGALS ASEVSKELNL VELKDRSWSI VASSAIKGEG ITEGLDWLID V IKEEQLAG ENLYFQSAGH HHHHH

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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

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 1.7 µm / Nominal defocus min: 1.3 µm
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 69.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: INSILICO MODEL / Details: PDB 5EE5 was also used as a startup model.
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 712548
FSC plot (resolution estimation)

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