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TitleMolecular mechanisms of inorganic-phosphate release from the core and barbed end of actin filaments.
Journal, issue, pagesNat Struct Mol Biol, Vol. 30, Issue 11, Page 1774-1785, Year 2023
Publish dateSep 25, 2023
AuthorsWout Oosterheert / Florian E C Blanc / Ankit Roy / Alexander Belyy / Micaela Boiero Sanders / Oliver Hofnagel / Gerhard Hummer / Peter Bieling / Stefan Raunser /
PubMed AbstractThe release of inorganic phosphate (P) from actin filaments constitutes a key step in their regulated turnover, which is fundamental to many cellular functions. The mechanisms underlying P release ...The release of inorganic phosphate (P) from actin filaments constitutes a key step in their regulated turnover, which is fundamental to many cellular functions. The mechanisms underlying P release from the core and barbed end of actin filaments remain unclear. Here, using human and bovine actin isoforms, we combine cryo-EM with molecular-dynamics simulations and in vitro reconstitution to demonstrate how actin releases P through a 'molecular backdoor'. While constantly open at the barbed end, the backdoor is predominantly closed in filament-core subunits and opens only transiently through concerted amino acid rearrangements. This explains why P escapes rapidly from the filament end but slowly from internal subunits. In a nemaline-myopathy-associated actin variant, the backdoor is predominantly open in filament-core subunits, resulting in accelerated P release and filaments with drastically shortened ADP-P caps. Our results provide the molecular basis for P release from actin and exemplify how a disease-linked mutation distorts the nucleotide-state distribution and atomic structure of the filament.
External linksNat Struct Mol Biol / PubMed:37749275 / PubMed Central
MethodsEM (single particle)
Resolution2.28 - 3.59 Å
Structure data

EMDB-16887, PDB-8oi6:
Cryo-EM structure of the undecorated barbed end of filamentous beta/gamma actin
Method: EM (single particle) / Resolution: 3.59 Å

EMDB-16888, PDB-8oi8:
Cryo-EM structure of ADP-bound, filamentous beta-actin harboring the R183W mutation
Method: EM (single particle) / Resolution: 2.28 Å

EMDB-16889, PDB-8oid:
Cryo-EM structure of ADP-bound, filamentous beta-actin harboring the N111S mutation
Method: EM (single particle) / Resolution: 2.3 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER / Water

Source
  • bos taurus (cattle)
  • amanita phalloides (death cap)
  • homo sapiens (human)
KeywordsSTRUCTURAL PROTEIN / Actin filament / cytoskeletal protein / ATPase

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