Irradiations of rabbit myofibrils with an ultraviolet microbeam. II. Phalloidin protects actin in solution but not in myofibrils from depolymerization by ultraviolet light.
We tested whether phalloidin protects actin in myofibrils from depolymerization by ultraviolet light (UV). I bands in glycerinated rabbit psoas myofibrils were irradiated with a UV microbeam in the presence and absence of phalloidin. We used the retention of contractility of the irradiated I band as the assay for protection of actin by phalloidin, since previous experiments indicated that UV blocks contraction of an irradiated I band by depolymerizing the thin filaments. The I bands of myofibrils incubated in phalloidin were as sensitive to UV as control I bands, indicating that phalloidin did not protect the thin filaments. However, phalloidin did protect F-actin in solution from depolymerization by UV. This apparent contradiction between F-actin in myofibrils and F-actin in solution was resolved by observing unirradiated myofibrils that were stained with rhodamine-phalloidin. It was found that phalloidin does not bind uniformly to the thin filaments, though as the fluorescence image is observed over time the staining pattern changes until it does appear to bind uniformly. We conclude that phalloidin does not protect F-actin in myofibrils from depolymerization by UV because it does not bind uniformly to the filaments.
D000199:Actins / Q000378:metabolism / Q000528:radiation effects*; D000818:Animals; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009132:Muscles / Q000187:drug effects / Q000528:radiation effects / Q000648:ultrastructure; D009210:Myofibrils / Q000187:drug effects / Q000528:radiation effects* / Q000648:ultrastructure; D009842:Oligopeptides / Q000494:pharmacology*; D010590:Phalloidine / Q000494:pharmacology*; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D014466:Ultraviolet Rays
D000199:Actins; D046911:Macromolecular Substances; D009842:Oligopeptides; D010590:Phalloidine