Structural insights into actin filament recognition by commonly used cellular actin markers
Autor: | Kutti R. Vinothkumar, Manjunath G Javoor, Minhajuddin Sirajuddin, Shubham Kesarwani, Archana Kumari |
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Rok vydání: | 2020 |
Předmět: |
Resource
cellular markers Models Molecular Conformational change actin cytoskeleton Cryo-electron microscopy Phalloidin macromolecular substances Computational biology Biology Filamentous actin General Biochemistry Genetics and Molecular Biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Structural Biology Utrophin Humans phalloidin Binding site Molecular Biology Actin 030304 developmental biology lifeAct 0303 health sciences General Immunology and Microbiology General Neuroscience Cryoelectron Microscopy Actin cytoskeleton Actins cryoEM chemistry Cell Adhesion Polarity & Cytoskeleton 030217 neurology & neurosurgery |
Zdroj: | The EMBO Journal |
ISSN: | 1460-2075 0261-4189 |
DOI: | 10.15252/embj.2019104006 |
Popis: | 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. Cryo‐EM structures of F‐actin complexed with phalloidin, LifeAct, or utrophin probes provide insights into structural determinants of actin marker binding and identify LifeAct preference for ADP‐bound actin. |
Databáze: | OpenAIRE |
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