+
Open data
-
Basic information
| Entry | ![]() | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Title | Ypt7 in complex with its GEF Mon1-Ccz1 | |||||||||
Map data | ||||||||||
Sample |
| |||||||||
Keywords | longin domain / GEF / nucleotide exchange / endosome / autophagosome / small GTPase / CYTOSOLIC PROTEIN | |||||||||
| Function / homology | Function and homology informationMon1-Ccz1 complex / regulation of vacuole fusion, non-autophagic / protein targeting to vacuole / multivesicular body membrane / fungal-type vacuole membrane / vesicle-mediated transport / autophagy / late endosome / GTPase activity / GTP binding Similarity search - Function | |||||||||
| Biological species | Thermochaetoides thermophila (fungus) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
Authors | Wilmes S / Schaefer J / Moeller A / Kuemmel D | |||||||||
| Funding support | Germany, 2 items
| |||||||||
Citation | Journal: Sci Adv / Year: 2025Title: Mechanistic adaptation of the metazoan RabGEFs Mon1-Ccz1 and Fuzzy-Inturned. Authors: Stephan Wilmes / Jesse Tönjes / Maik Drechsler / Anita Ruf / Jan-Hannes Schäfer / Anna Lürick / Dovile Januliene / Steven Apelt / Daniele Di Iorio / Seraphine V Wegner / Martin Loose / ...Authors: Stephan Wilmes / Jesse Tönjes / Maik Drechsler / Anita Ruf / Jan-Hannes Schäfer / Anna Lürick / Dovile Januliene / Steven Apelt / Daniele Di Iorio / Seraphine V Wegner / Martin Loose / Arne Moeller / Achim Paululat / Daniel Kümmel / ![]() Abstract: Rab GTPases organize intracellular trafficking and provide identity to organelles. Their spatiotemporal activation by guanine nucleotide exchange factors (GEFs) is tightly controlled to ensure ...Rab GTPases organize intracellular trafficking and provide identity to organelles. Their spatiotemporal activation by guanine nucleotide exchange factors (GEFs) is tightly controlled to ensure fidelity. Our structural and functional comparison of the tri-longin domain RabGEFs Mon1-Ccz1 and Fuzzy-Inturned reveals the molecular basis for their target specificity. Both complexes rely on a conserved sequence motif of their substrate GTPases for the catalytic mechanism, while secondary interactions allow discrimination between targets. We also find that dimeric Mon1-Ccz1 from fungi and the metazoan homologs with the additional third subunit RMC1/Bulli bind membranes through electrostatic interactions via distinct interfaces. Protein-lipid interaction studies and functional characterization in flies reveal an essential function of RMC1/Bulli as mediator of GEF complex membrane recruitment. In the case of Fuzzy-Inturned, reconstitution experiments demonstrate that the BAR (Bin-Amphiphysin-Rvs) domain protein CiBAR1 can support membrane recruitment of the GEF. Collectively, our study demonstrates the molecular basis for the adaptation of TLD-RabGEFs to different cellular functions. | |||||||||
| History |
|
-
Structure visualization
| Supplemental images |
|---|
-
Downloads & links
-EMDB archive
| Map data | emd_54212.map.gz | 161.5 MB | EMDB map data format | |
|---|---|---|---|---|
| Header (meta data) | emd-54212-v30.xml emd-54212.xml | 21 KB 21 KB | Display Display | EMDB header |
| Images | emd_54212.png | 38.7 KB | ||
| Filedesc metadata | emd-54212.cif.gz | 6.9 KB | ||
| Others | emd_54212_half_map_1.map.gz emd_54212_half_map_2.map.gz | 301.2 MB 301.2 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-54212 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-54212 | HTTPS FTP |
-Validation report
| Summary document | emd_54212_validation.pdf.gz | 828.9 KB | Display | EMDB validaton report |
|---|---|---|---|---|
| Full document | emd_54212_full_validation.pdf.gz | 828.5 KB | Display | |
| Data in XML | emd_54212_validation.xml.gz | 16.6 KB | Display | |
| Data in CIF | emd_54212_validation.cif.gz | 19.5 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-54212 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-54212 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9rs7MC ![]() 9rs6C ![]() 9rs8C ![]() 9rs9C M: atomic model generated by this map C: citing same article ( |
|---|---|
| Similar structure data | Similarity search - Function & homology F&H Search |
-
Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
|---|---|
| Related items in Molecule of the Month |
-
Map
| File | Download / File: emd_54212.map.gz / Format: CCP4 / Size: 325 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.68 Å | ||||||||||||||||||||||||||||||||||||
| Density |
| ||||||||||||||||||||||||||||||||||||
| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
|
-Supplemental data
-Half map: #2
| File | emd_54212_half_map_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Half map: #1
| File | emd_54212_half_map_2.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-
Sample components
-Entire : Complex of nucleotide free Ypt7 with its GEF Mon1-Ccz1
| Entire | Name: Complex of nucleotide free Ypt7 with its GEF Mon1-Ccz1 |
|---|---|
| Components |
|
-Supramolecule #1: Complex of nucleotide free Ypt7 with its GEF Mon1-Ccz1
| Supramolecule | Name: Complex of nucleotide free Ypt7 with its GEF Mon1-Ccz1 type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
|---|---|
| Source (natural) | Organism: Thermochaetoides thermophila (fungus) |
-Macromolecule #1: Rab small monomeric GTPase-like protein
| Macromolecule | Name: Rab small monomeric GTPase-like protein / type: protein_or_peptide / ID: 1 / Details: nucleotide free variant N125I / Number of copies: 1 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: Thermochaetoides thermophila (fungus) |
| Molecular weight | Theoretical: 23.218189 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: GSMSSRKKVL LKVIILGDSG VGKTSLMNQY VNKKFSASYK ATIGADFLTR EVMVDDRQVT MQLWDTAGQE RFQSLGVAFY RGADCCVLV FDVNNAKSFD ALDSWRDEFL IQASPRDPEN FPFVVLGIKI DVEESKRVIS TKRAQTFCQS KGGIPYFETS A KEAINVEE ...String: GSMSSRKKVL LKVIILGDSG VGKTSLMNQY VNKKFSASYK ATIGADFLTR EVMVDDRQVT MQLWDTAGQE RFQSLGVAFY RGADCCVLV FDVNNAKSFD ALDSWRDEFL IQASPRDPEN FPFVVLGIKI DVEESKRVIS TKRAQTFCQS KGGIPYFETS A KEAINVEE AFQVIARNAL MQEESEEFSG DFQDPINIHI ENERDGCAC UniProtKB: Rab small monomeric GTPase-like protein |
-Macromolecule #2: Vacuolar fusion protein MON1
| Macromolecule | Name: Vacuolar fusion protein MON1 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: Thermochaetoides thermophila (fungus) |
| Molecular weight | Theoretical: 72.198766 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: GPLGSMTQNN APESVAAEGT VAGQQTDTTK PEPSSQPAPE PPSQVTTDFP PNNKPNDVPA TEPASSSPRP TEPPAPPPPK PTIALSPLD IATLTFPDGT RGTFPAPPQT SAQSVSTPSI ASGHVTPQRD TDTASITSVA GTLRGVTGTA GDLASLLAGD G LGRKSKAW ...String: GPLGSMTQNN APESVAAEGT VAGQQTDTTK PEPSSQPAPE PPSQVTTDFP PNNKPNDVPA TEPASSSPRP TEPPAPPPPK PTIALSPLD IATLTFPDGT RGTFPAPPQT SAQSVSTPSI ASGHVTPQRD TDTASITSVA GTLRGVTGTA GDLASLLAGD G LGRKSKAW RVLRAQQQAC GEGSGEEGEI TEIEGLGLGE GMEGFERELD NIPDTLPDDE RLALWKGKLK HYLILSSAGK PI WSRHGDL SLVNSTMGVV QTIISFYEGA RNPLLGFTAG KVRFVILIKG PLYFVAISRL RESDAQLRAQ LEALYMQILS TLT LPILTN IFAHRPSTDL RGPLQGTESL LASLADSFTK GSPSTLLSAL ECLRLRKSQR QAITNIFLKS RCEELLYGLL VAGG KLVSV IRPRKHSLHP SDLQLIFNML FESGGIKGNG GENWIPLCLP AFNNTGYLYM YVSFLDDKAP DDQNQPPESS NLDAS NKNS SNTPDDDLTA LILISPSREA FYALQSMRTR LVSQLLSTGY LSLIRSTALS GRPSITSILP KTPLLHFLYK SRPNVQ WCM SSLSSLTPPG ATATETLLAR RKLMSVYEEL HAALHARHAH LRVVYSTADE KEGEGLACLG WSTPAFEVYC VAPGCVG RA GMAREVNRVV QWARREEERL FILGGGVF UniProtKB: Vacuolar fusion protein MON1 |
-Macromolecule #3: CCZ1/INTU/HSP4 first Longin domain-containing protein
| Macromolecule | Name: CCZ1/INTU/HSP4 first Longin domain-containing protein / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: Thermochaetoides thermophila (fungus) |
| Molecular weight | Theoretical: 75.867539 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: SMTTPVSPSP SGIIPAQLGF LAIYNPALGT TDETLEDQIV YYATASTLSQ ARRRHRRPRR RDRQRAQSVV KDSRPNAAGA TGDSEAVAE DKDPVSKEER HERLRQIGLA QGMVEFAKSF SDGEPVDTID TEKARVILVE VEEGWWILAS IDLTRLPLPQ I KTPTSSSA ...String: SMTTPVSPSP SGIIPAQLGF LAIYNPALGT TDETLEDQIV YYATASTLSQ ARRRHRRPRR RDRQRAQSVV KDSRPNAAGA TGDSEAVAE DKDPVSKEER HERLRQIGLA QGMVEFAKSF SDGEPVDTID TEKARVILVE VEEGWWILAS IDLTRLPLPQ I KTPTSSSA PPPAPNLNPL PPEPAYEYSS REVKPPSLLR ADLLRAYDLF LLHHGSSLSS LLASQGRAQL VASLTRFWDH FL ATWNVLL HGNPACDVFG GIKLAASGEL GIGVGEEERG SGEREVLEGL VERVEGLVDV VVGRYGGPPS EKGPEEEQWL GLG GEVGEE DGAVFLGVGA LDRKSLRGVV QWMEEVYVWG ENAFGKPRRD LSTGHFLLGL SECSEEELTS SQANPKAIFV ELKP SYQHP SRKIPPEDPQ PLGKVGPELP RDHTARLRPV IYVSQPFIYI LLFSEITPSP STWPTLAESL HAQLSPLQKP LLHST SYRP ERPVVETTSS SGTTTQHQIF DLVYDTETLT LQSTIPNIPD PFPYSATTPT GHSTGQQHHQ QSIWTRVEAL QTHAQI LAI LSSGRAIPTD PSSFTHLPWE EGERTCKTAR GWWIVWTRVV EHSPPDAVSL HHARDDDDND DDASCSVLGH LRSVSSS HA AGSTSSSSGS GFGLGAIPGL GGLGGWAADG ATRLAQGIGI DTRRYVEGLL TSLGR UniProtKB: CCZ1/INTU/HSP4 first Longin domain-containing protein, CCZ1/INTU/HSP4 first Longin domain-containing protein |
-Experimental details
-Structure determination
| Method | cryo EM |
|---|---|
Processing | single particle reconstruction |
| Aggregation state | particle |
-
Sample preparation
| Concentration | 0.8 mg/mL |
|---|---|
| Buffer | pH: 7.3 |
| Vitrification | Cryogen name: ETHANE |
-
Electron microscopy
| Microscope | TFS GLACIOS |
|---|---|
| Image recording | Film or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.8 µm |
Movie
Controller
About Yorodumi




Keywords
Thermochaetoides thermophila (fungus)
Authors
Germany, 2 items
Citation










Z (Sec.)
Y (Row.)
X (Col.)





































Processing
FIELD EMISSION GUN