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Yorodumi- EMDB-54631: Ternary cryo-EM structure of chicken ALG12 with Dol25-PP-GlcNAc2M... -
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Basic information
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| Title | Ternary cryo-EM structure of chicken ALG12 with Dol25-PP-GlcNAc2Man7, Dol25-P-Man, and Fab | |||||||||
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Keywords | Mannosyltransferase / ternary complex / N-linked glycosylation / TRANSFERASE | |||||||||
| Function / homology | Function and homology informationdol-P-Man:Man(7)GlcNAc(2)-PP-Dol alpha-1,6-mannosyltransferase activity / Biosynthesis of the N-glycan precursor (dolichol lipid-linked oligosaccharide, LLO) and transfer to a nascent protein / alpha-1,6-mannosyltransferase activity / dolichol-linked oligosaccharide biosynthetic process / protein N-linked glycosylation / Transferases; Glycosyltransferases; Hexosyltransferases / endoplasmic reticulum membrane Similarity search - Function | |||||||||
| Biological species | ![]() ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.42 Å | |||||||||
Authors | Alexander JAN / Chen SY / Mukherjee S / de Capitani M / Irobalieva RN / Rossi L / Agrawal P / Kowal J / Meirelles MA / Aebi M ...Alexander JAN / Chen SY / Mukherjee S / de Capitani M / Irobalieva RN / Rossi L / Agrawal P / Kowal J / Meirelles MA / Aebi M / Reymond JL / Kossiakoff AA / Riniker S / Locher KP | |||||||||
| Funding support | Switzerland, 2 items
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Citation | Journal: Nat Chem Biol / Year: 2026Title: Structures of ALG3/9/12 reveal the assembly logic of the N-glycan oligomannose core. Authors: J Andrew N Alexander / Shu-Yu Chen / Somnath Mukherjee / Mario de Capitani / Rossitza N Irobalieva / Lorenzo Rossi / Parth Agrawal / Julia Kowal / Matheus A Meirelles / Markus Aebi / Jean- ...Authors: J Andrew N Alexander / Shu-Yu Chen / Somnath Mukherjee / Mario de Capitani / Rossitza N Irobalieva / Lorenzo Rossi / Parth Agrawal / Julia Kowal / Matheus A Meirelles / Markus Aebi / Jean-Louis Reymond / Anthony A Kossiakoff / Sereina Riniker / Kaspar P Locher / ![]() Abstract: Asparagine-linked glycans are essential for the maturation and function of most eukaryotic secretory proteins. The biosynthesis and transfer of dolichylpyrophosphate-anchored GlcNAcManGlc glycan is a ...Asparagine-linked glycans are essential for the maturation and function of most eukaryotic secretory proteins. The biosynthesis and transfer of dolichylpyrophosphate-anchored GlcNAcManGlc glycan is a highly conserved process occurring in the endoplasmic reticulum (ER) membrane and involving over a dozen membrane proteins whose dysfunction is linked to congenital disorders of glycosylation (CDGs). Three membrane-integral mannosyltransferases, ALG3, ALG9 and ALG12, mediate four consecutive mannosylation reactions that convert GlcNAcMan to GlcNAcMan. Here, using chemoenzymatically synthesized lipid-linked glycan donor and acceptor analogs, we recapitulated this biosynthetic pathway in vitro. High-resolution cryo-electron microscopy structures of pseudo-Michaelis complexes of each step revealed how the branched glycan is accurately synthesized and unwanted side products are averted. Molecular dynamics simulations and mutagenesis studies uncovered a subtle but precise mechanism selecting the dolichylphosphomannose donor substrate over dolichylphosphoglucose, which is also present in the ER membrane. Our results also provide mechanistic explanations for enzyme dysfunction in CDGs and offer opportunities for N-glycan engineering. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_54631.map.gz | 483.2 MB | EMDB map data format | |
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| Header (meta data) | emd-54631-v30.xml emd-54631.xml | 24.6 KB 24.6 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_54631_fsc.xml | 16.9 KB | Display | FSC data file |
| Images | emd_54631.png | 56.1 KB | ||
| Filedesc metadata | emd-54631.cif.gz | 7.3 KB | ||
| Others | emd_54631_half_map_1.map.gz emd_54631_half_map_2.map.gz | 474.5 MB 474.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-54631 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-54631 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9s6tMC ![]() 9s6rC ![]() 9s6sC ![]() 9s6uC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_54631.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.65 Å | ||||||||||||||||||||||||||||||||||||
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #2
| File | emd_54631_half_map_1.map | ||||||||||||
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-Half map: #1
| File | emd_54631_half_map_2.map | ||||||||||||
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Sample components
+Entire : Ternary complex of chicken ALG12 with Gg12-11 Fab, anti-kappa lig...
+Supramolecule #1: Ternary complex of chicken ALG12 with Gg12-11 Fab, anti-kappa lig...
+Supramolecule #2: Anti-kapp light chain nanobody
+Supramolecule #3: Gg12-11 Fab
+Supramolecule #4: ALG12
+Macromolecule #1: Mannosyltransferase
+Macromolecule #2: Gg12-11 Fab heavy chain
+Macromolecule #3: Gg12-11 Fab light chain
+Macromolecule #4: [(3S,6Z,10E,14E)-3,7,11,15,19-pentamethylicosa-6,10,14,18-tetraen...
+Macromolecule #5: [(2~{R},3~{R},4~{R},5~{S},6~{R})-3-acetamido-5-[(2~{S},3~{R},4~{R...
+Macromolecule #6: CHOLESTEROL HEMISUCCINATE
+Macromolecule #7: Lauryl Maltose Neopentyl Glycol
+Macromolecule #8: water
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 3.9 mg/mL |
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| Buffer | pH: 7.5 |
| Vitrification | Cryogen name: ETHANE-PROPANE |
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Electron microscopy
| Microscope | TFS KRIOS |
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| Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 43.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 0.6 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi



Keywords

Authors
Switzerland, 2 items
Citation







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Homo sapiens (human)



Processing
FIELD EMISSION GUN

