Journal: EMBO J / Year: 2011 Title: Structural basis for the slow dynamics of the actin filament pointed end. Authors: Akihiro Narita / Toshiro Oda / Yuichiro Maéda / Abstract: The actin filament has clear polarity where one end, the pointed end, has a much slower polymerization and depolymerization rate than the other end, the barbed end. This intrinsic difference of the ...The actin filament has clear polarity where one end, the pointed end, has a much slower polymerization and depolymerization rate than the other end, the barbed end. This intrinsic difference of the ends significantly affects all actin dynamics in the cell, which has central roles in a wide spectrum of cellular functions. The detailed mechanism underlying this difference has remained elusive, because high-resolution structures of the filament ends have not been available. Here, we present the structure of the actin filament pointed end obtained using a single particle analysis of cryo-electron micrographs. We determined that the terminal pointed end subunit is tilted towards the penultimate subunit, allowing specific and extra loop-to-loop inter-strand contacts between the two end subunits, which is not possible in other parts of the filament. These specific contacts prevent the end subunit from dissociating. For elongation, the loop-to-loop contacts also inhibit the incorporation of another actin monomer at the pointed end. These observations are likely to account for the less dynamic pointed end.
History
Deposition
Feb 2, 2011
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Header (metadata) release
Feb 9, 2011
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Map release
Mar 25, 2011
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Update
Mar 13, 2013
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Current status
Mar 13, 2013
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
pH: 7.4 Details: 50 mM NaCl, 10 mM sodium phosphate buffer (pH 7.4), 3 mM MgCl2, 0.005% (w/v) NaN3, 0.7 mM DTT
Grid
Details: quantifoil R2/2
Vitrification
Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 4 K / Instrument: OTHER / Method: Blot for 3 seconds before plunging
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Electron microscopy
Microscope
JEOL 3200FSC
Temperature
Average: 4 K
Specialist optics
Energy filter - Name: JEOL / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV
Image recording
Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: ZEISS SCAI / Digitization - Sampling interval: 7 µm / Number real images: 142 / Average electron dose: 34 e/Å2 / Bits/pixel: 12
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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