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Title | Structural insights into actin filament recognition by commonly used cellular actin markers. |
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Journal, issue, pages | EMBO J, Vol. 39, Issue 14, Page e104006, Year 2020 |
Publish date | Jul 15, 2020 |
Authors | Archana Kumari / Shubham Kesarwani / Manjunath G Javoor / Kutti R Vinothkumar / Minhajuddin Sirajuddin / |
PubMed 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 ...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. |
External links | EMBO J / PubMed:32567727 / PubMed Central |
Methods | EM (helical sym.) |
Resolution | 3.6 - 4.2 Å |
Structure data | EMDB-30085, PDB-6m5g: EMDB-30171: F-actin-ADP-state EMDB-30177, PDB-7bte: EMDB-30179, PDB-7bti: |
Chemicals | ChemComp-MG: ChemComp-ADP: |
Source |
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Keywords | CONTRACTILE PROTEIN / Utrophin / N-terminus actin binding domain / calponin homology / F-actin / F-actin marker protein / ADP-F-actin / CONTRACTILE PROTEIN/PROTEIN BINDING / CONTRACTILE PROTEIN-PROTEIN BINDING complex |