+
Open data
-
Basic information
Entry | ![]() | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | Low resolution 3D reconstruction of ATG2A from cryo-EM | ||||||||||||
![]() | |||||||||||||
![]() |
| ||||||||||||
![]() | Lipid transfer protein / autophagy / LIPID TRANSPORT | ||||||||||||
Function / homology | ![]() organelle membrane contact site / phagophore / lipid transfer activity / glycophagy / positive regulation of autophagosome assembly / phagophore assembly site membrane / piecemeal microautophagy of the nucleus / phosphatidylinositol-3-phosphate binding / phagophore assembly site / reticulophagy ...organelle membrane contact site / phagophore / lipid transfer activity / glycophagy / positive regulation of autophagosome assembly / phagophore assembly site membrane / piecemeal microautophagy of the nucleus / phosphatidylinositol-3-phosphate binding / phagophore assembly site / reticulophagy / autophagosome assembly / protein-membrane adaptor activity / lipid droplet / endoplasmic reticulum membrane Similarity search - Function | ||||||||||||
Biological species | ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 14.7 Å | ||||||||||||
![]() | Cherepanov P / Chiduza GN / Pye VE / van Vliet AR / Tooze SA | ||||||||||||
Funding support | ![]()
| ||||||||||||
![]() | ![]() Title: ATG9A and ATG2A form a heteromeric complex essential for autophagosome formation. Authors: Alexander R van Vliet / George N Chiduza / Sarah L Maslen / Valerie E Pye / Dhira Joshi / Stefano De Tito / Harold B J Jefferies / Evangelos Christodoulou / Chloë Roustan / Emma Punch / ...Authors: Alexander R van Vliet / George N Chiduza / Sarah L Maslen / Valerie E Pye / Dhira Joshi / Stefano De Tito / Harold B J Jefferies / Evangelos Christodoulou / Chloë Roustan / Emma Punch / Javier H Hervás / Nicola O'Reilly / J Mark Skehel / Peter Cherepanov / Sharon A Tooze / ![]() ![]() Abstract: ATG9A and ATG2A are essential core members of the autophagy machinery. ATG9A is a lipid scramblase that allows equilibration of lipids across a membrane bilayer, whereas ATG2A facilitates lipid flow ...ATG9A and ATG2A are essential core members of the autophagy machinery. ATG9A is a lipid scramblase that allows equilibration of lipids across a membrane bilayer, whereas ATG2A facilitates lipid flow between tethered membranes. Although both have been functionally linked during the formation of autophagosomes, the molecular details and consequences of their interaction remain unclear. By combining data from peptide arrays, crosslinking, and hydrogen-deuterium exchange mass spectrometry together with cryoelectron microscopy, we propose a molecular model of the ATG9A-2A complex. Using this integrative structure modeling approach, we identify several interfaces mediating ATG9A-2A interaction that would allow a direct transfer of lipids from ATG2A into the lipid-binding perpendicular branch of ATG9A. Mutational analyses combined with functional activity assays demonstrate their importance for autophagy, thereby shedding light on this protein complex at the heart of autophagy. | ||||||||||||
History |
|
-
Structure visualization
Supplemental images |
---|
-
Downloads & links
-EMDB archive
Map data | ![]() | 1.2 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 16.9 KB 16.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 5.1 KB | Display | ![]() |
Images | ![]() | 75.1 KB | ||
Masks | ![]() | 10.5 MB | ![]() | |
Filedesc metadata | ![]() | 6.2 KB | ||
Others | ![]() ![]() | 7.9 MB 7.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 187.5 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 187.1 KB | Display | |
Data in XML | ![]() | 503 B | Display | |
Data in CIF | ![]() | 373 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article ( |
---|---|
Similar structure data | Similarity search - Function & homology ![]() |
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.52 Å | ||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Mask #1
File | ![]() | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #1
File | emd_15605_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #2
File | emd_15605_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-
Sample components
-Entire : Purified ATG2A
Entire | Name: Purified ATG2A |
---|---|
Components |
|
-Supramolecule #1: Purified ATG2A
Supramolecule | Name: Purified ATG2A / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
---|---|
Source (natural) | Organism: ![]() |
-Macromolecule #1: Autophagy-related protein 2 homolog A
Macromolecule | Name: Autophagy-related protein 2 homolog A / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
---|---|
Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: MDYKDDDDKD YKDDDDKDYK DDDDKGSGAG AGAGAILNSR VSGLRSRMSR WLWPWSNCVK ERVCRYLLHH YLGHFFQEHL SLDQLSLDLY KGSVALRDIH LEIWSVNEVL ESMESPLELV EGFVGSIEVA VPWAALLTDH CTVRVSGLQL TLQPRRGPAP GAADSQSWAS ...String: MDYKDDDDKD YKDDDDKDYK DDDDKGSGAG AGAGAILNSR VSGLRSRMSR WLWPWSNCVK ERVCRYLLHH YLGHFFQEHL SLDQLSLDLY KGSVALRDIH LEIWSVNEVL ESMESPLELV EGFVGSIEVA VPWAALLTDH CTVRVSGLQL TLQPRRGPAP GAADSQSWAS CMTTSLQLAQ ECLRDGLPEP SEPPQPLEGL EMFAQTIETV LRRIKVTFLD TVVRVEHSPG DGERGVAVEV RVQRLEYCDE AVRDPSQAPP VDVHQPPAFL HKLLQLAGVR LHYEELPAQE EPPEPPLQIG SCSGYMELMV KLKQNEAFPG PKLEVAGQLG SLHLLLTPRQ LQQLQELLSA VSLTDHEGLA DKLNKSRPLG AEDLWLIEQD LNQQLQAGAV AEPLSPDPLT NPLLNLDNTD LFFSMAGLTS SVASALSELS LSDVDLASSV RSDMASRRLS AQAHPAGKMA PNPLLDTMRP DSLLKMTLGG VTLTLLQTSA PSSGPPDLAT HFFTEFDATK DGPFGSRDFH HLRPRFQRAC PCSHVRLTGT AVQLSWELRT GSRGRRTTSM EVHFGQLEVL ECLWPRGTSE PEYTEILTFP GTLGSQASAR PCAHLRHTQI LRRVPKSRPR RSVACHCHSE LALDLANFQA DVELGALDRL AALLRLATVP AEPPAGLLTE PLPAMEQQTV FRLSAPRATL RLRFPIADLR PEPDPWAGQA VRAEQLRLEL SEPQFRSELS SGPGPPVPTH LELTCSDLHG IYEDGGKPPV PCLRVSKALD PKSTGRKYFL PQVVVTVNPQ SSSTQWEVAP EKGEELELSV ESPCELREPE PSPFSSKRTM YETEEMVIPG DPEEMRTFQS RTLALSRCSL EVILPSVHIF LPSKEVYESI YNRINNDLLM WEPADLLPTP DPAAQPSGFP GPSGFWHDSF KMCKSAFKLA NCFDLTPDSD SDDEDAHFFS VGASGGPQAA APEAPSLHLQ STFSTLVTVL KGRITALCET KDEGGKRLEA VHGELVLDME HGTLFSVSQY CGQPGLGYFC LEAEKATLYH RAAVDDYPLP SHLDLPSFAP PAQLAPTIYP SEEGVTERGA SGRKGQGRGP HMLSTAVRIH LDPHKNVKEF LVTLRLHKAT LRHYMALPEQ SWHSQLLEFL DVLDDPVLGY LPPTVITILH THLFSCSVDY RPLYLPVRVL ITAETFTLSS NIIMDTSTFL LRFILDDSAL YLSDKCEVET LDLRRDYVCV LDVDLLELVI KTWKGSTEGK LSQPLFELRC SNNVVHVHSC ADSCALLVNL LQYVMSTGDL HPPPRPPSPT EIAGQKLSES PASLPSCPPV ETALINQRDL ADALLDTERS LRELAQPSGG HLPQASPISV YLFPGERSGA PPPSPPVGGP AGSLGSCSEE KEDEREEEGD GDTLDSDEFC ILDAPGLGIP PRDGEPVVTQ LHPGPIVVRD GYFSRPIGST DLLRAPAHFP VPSTRVVLRE VSLVWHLYGG RDFGPHPGHR ARTGLSGPRS SPSRCSGPNR PQNSWRTQGG SGRQHHVLME IQLSKVSFQH EVYPAEPATG PAAPSQELEE RPLSRQVFIV QELEVRDRLA SSQINKFLYL HTSERMPRRA HSNMLTIKAL HVAPTTNLGG PECCLRVSLM PLRLNVDQDA LFFLKDFFTS LVAGINPVVP GETSAEARPE TRAQPSSPLE GQAEGVETTG SQEAPGGGHS PSPPDQQPIY FREFRFTSEV PIWLDYHGKH VTMDQVGTFA GLLIGLAQLN CSELKLKRLC CRHGLLGVDK VLGYALNEWL QDIRKNQLPG LLGGVGPMHS VVQLFQGFRD LLWLPIEQYR KDGRLMRGLQ RGAASFGSST ASAALELSNR LVQAIQATAE TVYDILSPAA PVSRSLQDKR SARRLRRGQQ PADLREGVAK AYDTVREGIL DTAQTICDVA SRGHEQKGLT GAVGGVIRQL PPTVVKPLIL ATEATSSLLG GMRNQIVPDA HKDHALKWRS DSAQDNSAVD GTAGPGSTGS R UniProtKB: Autophagy-related protein 2 homolog A |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Concentration | 0.56 mg/mL |
---|---|
Buffer | pH: 8 / Details: 50 mM HEPES, 200 mM NaCl, 1 mM TCEP |
Grid | Model: C-flat-1.2/1.3 / Material: GOLD / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295.15 K / Instrument: FEI VITROBOT MARK IV |
Details | Grids prepared straight after size exclusion elution. |
-
Electron microscopy
Microscope | FEI TALOS ARCTICA |
---|---|
Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 8.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.0 µm |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |