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Structural insights into actin filament recognition by commonly used cellular actin markers
Author(s) -
Kumari Archana,
Kesarwani Shubham,
Javoor Manjunath G,
Vinothkumar Kutti R,
Sirajuddin Minhajuddin
Publication year - 2020
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.2019104006
Subject(s) - actin , biology , phalloidin , actin binding protein , microbiology and biotechnology , cytoskeleton , actin remodeling , actin cytoskeleton , utrophin , biophysics , biochemistry , cell
Abstract 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.