7K21
Cryo-EM structure of pyrene-labeled ADP-Pi-actin filaments
Summary for 7K21
| Entry DOI | 10.2210/pdb7k21/pdb |
| EMDB information | 22639 |
| Descriptor | Actin, alpha skeletal muscle, MAGNESIUM ION, ADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total) |
| Functional Keywords | actin, pyrene, fluorescence, adp, phosphate, cytosolic protein |
| Biological source | Gallus gallus (Chicken) |
| Total number of polymer chains | 4 |
| Total formula weight | 170725.65 |
| Authors | Chou, S.Z.,Pollard, T.D. (deposition date: 2020-09-08, release date: 2020-11-04, Last modification date: 2025-04-02) |
| Primary citation | 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 Cited by 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: 33214556DOI: 10.1038/s41467-020-19762-1 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3 Å) |
Structure validation
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