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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-14066 | |||||||||
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Title | Structure of the Rab GEF complex Mon1-Ccz1 | |||||||||
![]() | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Map sharpened by Phenix local anisotropic filtering. | |||||||||
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![]() | guanine nucleotide exchange factor / TLD Rab GEF / longin domains / PIP binding / ENDOCYTOSIS | |||||||||
Function / homology | ![]() Mon1-Ccz1 complex / protein targeting to vacuole / multivesicular body membrane / fungal-type vacuole membrane / vesicle-mediated transport / autophagy Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.85 Å | |||||||||
![]() | Klink BU / Herrmann E | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure of the Mon1-Ccz1 complex reveals molecular basis of membrane binding for Rab7 activation. Authors: Björn U Klink / Eric Herrmann / Claudia Antoni / Lars Langemeyer / Stephan Kiontke / Christos Gatsogiannis / Christian Ungermann / Stefan Raunser / Daniel Kümmel / ![]() Abstract: Activation of the GTPase Rab7/Ypt7 by its cognate guanine nucleotide exchange factor (GEF) Mon1-Ccz1 marks organelles such as endosomes and autophagosomes for fusion with lysosomes/vacuoles and ...Activation of the GTPase Rab7/Ypt7 by its cognate guanine nucleotide exchange factor (GEF) Mon1-Ccz1 marks organelles such as endosomes and autophagosomes for fusion with lysosomes/vacuoles and degradation of their content. Here, we present a high-resolution cryogenic electron microscopy structure of the Mon1-Ccz1 complex that reveals its architecture in atomic detail. Mon1 and Ccz1 are arranged side by side in a pseudo-twofold symmetrical heterodimer. The three Longin domains of each Mon1 and Ccz1 are triangularly arranged, providing a strong scaffold for the catalytic center of the GEF. At the opposite side of the Ypt7-binding site, a positively charged and relatively flat patch stretches the Longin domains 2/3 of Mon1 and functions as a phosphatidylinositol phosphate-binding site, explaining how the GEF is targeted to membranes. Our work provides molecular insight into the mechanisms of endosomal Rab activation and serves as a blueprint for understanding the function of members of the Tri Longin domain Rab-GEF family. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 20.4 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.8 KB 23.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 8.3 KB | Display | ![]() |
Images | ![]() | 77.3 KB | ||
Filedesc metadata | ![]() | 6.9 KB | ||
Others | ![]() ![]() ![]() ![]() | 19.2 MB 20.8 MB 17 MB 17 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7qlaMC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Map sharpened by Phenix local anisotropic filtering. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.09 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665;...
File | emd_14066_additional_1.map | ||||||||||||
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Annotation | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Map created by DeepEMhancer with high resolution training model. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665;...
File | emd_14066_additional_2.map | ||||||||||||
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Annotation | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Map created by Relion Postprocessing. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665;...
File | emd_14066_half_map_1.map | ||||||||||||
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Annotation | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Unfiltered half-map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665;...
File | emd_14066_half_map_2.map | ||||||||||||
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Annotation | Cryo-EM structure of the Mon1-Ccz1 complex (CtMon1 aa141-665; CtCcz1 aa1-796delta361-460). Unfiltered half-map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Rab GEF complex Mon1-Ccz1
Entire | Name: Rab GEF complex Mon1-Ccz1 |
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Components |
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-Supramolecule #1: Rab GEF complex Mon1-Ccz1
Supramolecule | Name: Rab GEF complex Mon1-Ccz1 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all / Details: CtMon1 aa141-665; CtCcz1 aa1-796delta361-460 |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Vacuolar fusion protein MON1
Macromolecule | Name: Vacuolar fusion protein MON1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 57.571832 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: TGTAGDLASL LAGDGLGRKS KAWRVLRAQQ QACGEGSGEE GEITEIEGLG LGEGMEGFER ELDNIPDTLP DDERLALWKG KLKHYLILS SAGKPIWSRH GDLSLVNSTM GVVQTIISFY EGARNPLLGF TAGKVRFVIL IKGPLYFVAI SRLRESDAQL R AQLEALYM ...String: TGTAGDLASL LAGDGLGRKS KAWRVLRAQQ QACGEGSGEE GEITEIEGLG LGEGMEGFER ELDNIPDTLP DDERLALWKG KLKHYLILS SAGKPIWSRH GDLSLVNSTM GVVQTIISFY EGARNPLLGF TAGKVRFVIL IKGPLYFVAI SRLRESDAQL R AQLEALYM QILSTLTLPI LTNIFAHRPS TDLRGPLQGT ESLLASLADS FTKGSPSTLL SALECLRLRK SQRQAITNIF LK SRCEELL YGLLVAGGKL VSVIRPRKHS LHPSDLQLIF NMLFESGGIK GNGGENWIPL CLPAFNNTGY LYMYVSFLDD KAP DDQNQP PESSNLDASN KNSSNTPDDD LTALILISPS REAFYALQSM RTRLVSQLLS TGYLSLIRST ALSGRPSITS ILPK TPLLH FLYKSRPNVQ WCMSSLSSLT PPGATATETL LARRKLMSVY EELHAALHAR HAHLRVVYST ADEKEGEGLA CLGWS TPAF EVYCVAPGCV GRAGMAREVN RVVQWARREE ERLFILGGGV F UniProtKB: Vacuolar fusion protein MON1 |
-Macromolecule #2: Ccz1
Macromolecule | Name: Ccz1 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 75.780461 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MTTPVSPSPS GIIPAQLGFL AIYNPALGTT DETLEDQIVY YATASTLSQA RRRHRRPRRR DRQRAQSVVK DSRPNAAGAT GDSEAVAED KDPVSKEERH ERLRQIGLAQ GMVEFAKSFS DGEPVDTIDT EKARVILVEV EEGWWILASI DLTRLPLPQI K TPTSSSAP ...String: MTTPVSPSPS GIIPAQLGFL AIYNPALGTT DETLEDQIVY YATASTLSQA RRRHRRPRRR DRQRAQSVVK DSRPNAAGAT GDSEAVAED KDPVSKEERH ERLRQIGLAQ GMVEFAKSFS DGEPVDTIDT EKARVILVEV EEGWWILASI DLTRLPLPQI K TPTSSSAP PPAPNLNPLP PEPAYEYSSR EVKPPSLLRA DLLRAYDLFL LHHGSSLSSL LASQGRAQLV ASLTRFWDHF LA TWNVLLH GNPACDVFGG IKLAASGELG IGVGEEERGS GEREVLEGLV ERVEGLVDVV VGRYGGPPSE KGPEEEQWLG LGG EVGEED GAVFLGVGAL DRKSLRGVVQ WMEEVYVWGE NAFGKPRRDL STGHFLLGLS ECSEEELTSS QANPKAIFVE LKPS YQHPS RKIPPEDPQP LGKVGPELPR DHTARLRPVI YVSQPFIYIL LFSEITPSPS TWPTLAESLH AQLSPLQKPL LHSTS YRPE RPVVETTSSS GTTTQHQIFD LVYDTETLTL QSTIPNIPDP FPYSATTPTG HSTGQQHHQQ SIWTRVEALQ THAQIL AIL SSGRAIPTDP SSFTHLPWEE GERTCKTARG WWIVWTRVVE HSPPDAVSLH HARDDDDNDD DASCSVLGHL RSVSSSH AA GSTSSSSGSG FGLGAIPGLG GLGGWAADGA TRLAQGIGID TRRYVEGLLT SLGR |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.3 Component:
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Grid | Model: Quantifoil R2/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK II |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 11916 / Average exposure time: 15.0 sec. / Average electron dose: 73.0 e/Å2 Details: Images were collected in movie-mode with 4 frames per second |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 0.001 mm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Protocol: AB INITIO MODEL |
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Output model | ![]() PDB-7qla: |