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TitleArchitecture of the AP2/clathrin coat on the membranes of clathrin-coated vesicles.
Journal, issue, pagesSci Adv, Vol. 6, Issue 30, Page eaba8381, Year 2020
Publish dateJul 22, 2020
AuthorsOleksiy Kovtun / Veronica Kane Dickson / Bernard T Kelly / David J Owen / John A G Briggs /
PubMed AbstractClathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which ...Clathrin-mediated endocytosis (CME) is crucial for modulating the protein composition of a cell's plasma membrane. Clathrin forms a cage-like, polyhedral outer scaffold around a vesicle, to which cargo-selecting clathrin adaptors are attached. Adaptor protein complex (AP2) is the key adaptor in CME. Crystallography has shown AP2 to adopt a range of conformations. Here, we used cryo-electron microscopy, tomography, and subtomogram averaging to determine structures, interactions, and arrangements of clathrin and AP2 at the key steps of coat assembly, from AP2 in solution to membrane-assembled clathrin-coated vesicles (CCVs). AP2 binds cargo and PtdIns(4,5) (phosphatidylinositol 4,5-bisphosphate)-containing membranes via multiple interfaces, undergoing conformational rearrangement from its cytosolic state. The binding mode of AP2 β2 appendage into the clathrin lattice in CCVs and buds implies how the adaptor structurally modulates coat curvature and coat disassembly.
External linksSci Adv / PubMed:32743075 / PubMed Central
MethodsEM (single particle) / EM (subtomogram averaging)
Resolution3.9 - 11.9 Å
Structure data

10747

6yae

EMDB-10747, PDB-6yae:
AP2 core in physiological buffer
Method: EM (single particle) / Resolution: 3.9 Å

10748

6yaf

EMDB-10748, PDB-6yaf:
AP2 on a membrane containing tyrosine-based cargo peptide
Method: EM (subtomogram averaging) / Resolution: 9.1 Å

EMDB-10749:
AP2 dimer on a membrane containing tyrosine-based cargo peptide
Method: EM (subtomogram averaging) / Resolution: 10.2 Å

EMDB-10750:
AP2 in clathrin coats assembled on a membrane containing tyrosine-based cargo peptide
Method: EM (subtomogram averaging) / Resolution: 11.9 Å

10751

6yah

EMDB-10751, PDB-6yah:
AP2 in clathrin coats assembled on a membrane containing dileucine- and tyrosine-based cargo peptides
Method: EM (subtomogram averaging) / Resolution: 10.2 Å

EMDB-10752:
Hexagon bounding clathrin leg in clathrin coats assembled on a membrane
Method: EM (subtomogram averaging) / Resolution: 7.7 Å

EMDB-10753:
Pentagon bounding clathrin legs in clathrin coats assembled on a membrane
Method: EM (subtomogram averaging) / Resolution: 7.7 Å

10754

6yai

EMDB-10754, PDB-6yai:
Clathrin with bound beta2 appendage of AP2
Method: EM (subtomogram averaging) / Resolution: 9.2 Å

Source
  • rattus norvegicus (Norway rat)
  • homo sapiens (human)
  • mus musculus (house mouse)
  • sus scrofa (pig)
KeywordsENDOCYTOSIS / clathrin / clathrin adaptor / ap2 / clathrin assembly

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