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TitleStructural Effects and Functional Implications of Phalloidin and Jasplakinolide Binding to Actin Filaments.
Journal, issue, pagesStructure, Vol. 28, Issue 4, Page 437-449.e5, Year 2020
Publish dateApr 7, 2020
AuthorsSabrina Pospich / Felipe Merino / Stefan Raunser /
PubMed AbstractActin undergoes structural transitions during polymerization, ATP hydrolysis, and subsequent release of inorganic phosphate. Several actin-binding proteins sense specific states during this ...Actin undergoes structural transitions during polymerization, ATP hydrolysis, and subsequent release of inorganic phosphate. Several actin-binding proteins sense specific states during this transition and can thus target different regions of the actin filament. Here, we show in atomic detail that phalloidin, a mushroom toxin that is routinely used to stabilize and label actin filaments, suspends the structural changes in actin, likely influencing its interaction with actin-binding proteins. Furthermore, high-resolution cryoelectron microscopy structures reveal structural rearrangements in F-actin upon inorganic phosphate release in phalloidin-stabilized filaments. We find that the effect of the sponge toxin jasplakinolide differs from the one of phalloidin, despite their overlapping binding site and similar interactions with the actin filament. Analysis of structural conformations of F-actin suggests that stabilizing agents trap states within the natural conformational space of actin.
External linksStructure / PubMed:32084355
MethodsEM (single particle)
Resolution3.1 - 3.7 Å
Structure data

10363

6t1y

EMDB-10363, PDB-6t1y:
Cryo-EM structure of phalloidin-stabilized F-actin (copolymerized)
Method: EM (single particle) / Resolution: 3.3 Å

10364

6t20

EMDB-10364, PDB-6t20:
Cryo-EM structure of phalloidin-stabilized F-actin (aged)
Method: EM (single particle) / Resolution: 3.7 Å

10365

6t23

EMDB-10365, PDB-6t23:
Cryo-EM structure of jasplakinolide-stabilized F-actin (aged)
Method: EM (single particle) / Resolution: 3.1 Å

10366

6t24

EMDB-10366, PDB-6t24:
Cryo-EM structure of jasplakinolide-stabilized F-actin (aged)
Method: EM (single particle) / Resolution: 3.7 Å

10367

6t25

EMDB-10367, PDB-6t25:
Cryo-EM structure of phalloidin-Alexa Flour-546-stabilized F-actin (copolymerized)
Method: EM (single particle) / Resolution: 3.6 Å

Chemicals

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

ChemComp-MG:
Unknown entry

ChemComp-PO4:
PHOSPHATE ION / Phosphate

ChemComp-9ZK:
(4~{R},7~{R},10~{S},13~{S},15~{E},19~{S})-10-(4-azanylbutyl)-4-(4-hydroxyphenyl)-7-(1~{H}-indol-3-ylmethyl)-8,13,15,19-tetramethyl-1-oxa-5,8,11-triazacyclononadec-15-ene-2,6,9,12-tetrone

Source
  • oryctolagus cuniculus (rabbit)
  • synthetic construct (others)
  • Rabbit (rabbit)
KeywordsSTRUCTURAL PROTEIN / Cytoskeleton / phalloidin / stabilized-actin filament / jasplakinolide / stabilized actin filament

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