7K20

Cryo-EM structure of pyrene-labeled ADP-actin filaments

7K20 の概要

• エントリーDOI: 10.2210/pdb7k20/pdb
• EMDBエントリー: 22638
• 分子名称: Actin, alpha skeletal muscle, MAGNESIUM ION, ADENOSINE-5'-DIPHOSPHATE, ... (4 entities in total)
• 機能のキーワード: actin, pyrene, fluorescence, adp, cytosolic protein
• 由来する生物種: Gallus gallus (Chicken)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 170345.76

構造登録者

Chou, S.Z.,Pollard, T.D. (登録日: 2020-09-08, 公開日: 2020-11-04, 最終更新日: 2025-04-02)

主引用文献

Chou, S.Z.,Pollard, T.D.
Cryo-electron microscopy structures of pyrene-labeled ADP-P i - and ADP-actin filaments.
Nat Commun, 11:5897-5897, 2020

PubMed Abstract: Since the fluorescent reagent N-(1-pyrene)iodoacetamide was first used to label skeletal muscle actin in 1981, the pyrene-labeled actin has become the most widely employed tool to measure the kinetics of actin polymerization and the interaction between actin and actin-binding proteins. Here we report high-resolution cryo-electron microscopy structures of actin filaments with N-1-pyrene conjugated to cysteine 374 and either ADP (3.2 Å) or ADP-phosphate (3.0 Å) in the active site. Polymerization buries pyrene in a hydrophobic cavity between subunits along the long-pitch helix with only minor differences in conformation compared with native actin filaments. These structures explain how polymerization increases the fluorescence 20-fold, how myosin and cofilin binding to filaments reduces the fluorescence, and how profilin binding to actin monomers increases the fluorescence.

PubMed: 33214556
DOI: 10.1038/s41467-020-19762-1

実験手法

ELECTRON MICROSCOPY (3.2 Å)