%0 Journal Article %J EMBO J %D 2020 %T Structural insights into actin filament recognition by commonly used cellular actin markers. %A Kumari, Archana %A Kesarwani, Shubham %A Javoor, Manjunath G %A Vinothkumar, Kutti R %A Sirajuddin, Minhajuddin %X

Cellular studies of filamentous actin (F-actin) processes commonly utilize fluorescent versions of toxins, peptides, and proteins that bind actin. While the choice of these markers has been largely based on availability and ease, there is a severe dearth of structural data for an informed judgment in employing suitable F-actin markers for a particular requirement. Here, we describe the electron cryomicroscopy structures of phalloidin, lifeAct, and utrophin bound to F-actin, providing a comprehensive high-resolution structural comparison of widely used actin markers and their influence towards F-actin. Our results show that phalloidin binding does not induce specific conformational change and lifeAct specifically recognizes closed D-loop conformation, i.e., ADP-Pi or ADP states of F-actin. The structural models aided designing of minimal utrophin and a shorter lifeAct, which can be utilized as F-actin marker. Together, our study provides a structural perspective, where the binding sites of utrophin and lifeAct overlap with majority of actin-binding proteins and thus offering an invaluable resource for researchers in choosing appropriate actin markers and generating new marker variants.

%B EMBO J %V 39 %P e104006 %8 2020 Jul 15 %G eng %N 14 %R 10.15252/embj.2019104006