Conformation changes of actin during formation of filaments and paracrystals and upon interaction with DNase I, cytochalasin B, and phalloidin.
Spin labels attached to rabbit muscle actin became more immobilized upon conversion of actin from the G state to the F state with 50 mM KCl. Titration of G-actin with MgCl2 produced F-actin-like EPR spectra between 2 and 5 mM-actin filaments by electron microscopy. Higher concentrations of MgCl2 produced bundles of actin and eventually paracrystals, accompanied by further immobilization of spin labels. The effects of MgCl2 and KCl were competitive: addition of MgCl2 to 50 mM could convert F-actin (50 mM KCl) to paracrystalline (P) actin; the reverse titration (0 to 200 mM KCl in the presence of 20 mM MgCl2) was less complete. Addition of DNase I to G- or F-actin gave the expected amorphous electron micrographic pattern, and the actin was not sedimentable at (400,000 x g x h). EPR showed that the actin was in the G conformation. Addition of DNase I to paracrystalline actin gave the F conformation (EPR) but the actin was "G" by electron microscopy. Phalloidin converted G-actin to F-actin, had no effect on F-actin, and converted P-actin to the F state by electron microscopy but maintained the P conformation by EPR. Cytochalasin B produced no effects observable by EPR or centrifugation but "untwisted" paracrystals into nets. Since actin retained its P conformation by EPR in two states which were morphologically not P, we conclude that the P state is a distinct conformation of the actin molecule and that actin filaments aggregate to form bundles (and eventually paracrystals) when actin monomers are able to enter the P conformation.
D000199:Actins; D000818:Animals; D003460:Crystallization; D003571:Cytochalasin B; D003851:Deoxyribonucleases; D004578:Electron Spin Resonance Spectroscopy; D004720:Endonucleases; D046911:Macromolecular Substances; D008274:Magnesium; D008854:Microscopy, Electron; D008958:Models, Molecular; D008968:Molecular Conformation; D009132:Muscles / Q000032:analysis; D009842:Oligopeptides; D010590:Phalloidine; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D013113:Spin Labels
D000199:Actins; D046911:Macromolecular Substances; D009842:Oligopeptides; D013113:Spin Labels; D010590:Phalloidine; D003571:Cytochalasin B; D003851:Deoxyribonucleases; D004720:Endonucleases; D008274:Magnesium