1. New insights into the function of Fascin in actin bundling: A combined theoretical and experimental study.
- Author
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Wu X, Wen B, Lin L, Shi W, Li D, Cheng Y, Xu LY, Li EM, and Dong G
- Subjects
- Humans, Mutation, Protein Binding, Animals, Actin Cytoskeleton metabolism, Actin Cytoskeleton genetics, Phosphorylation, Microfilament Proteins metabolism, Microfilament Proteins genetics, Microfilament Proteins chemistry, Carrier Proteins metabolism, Carrier Proteins genetics, Carrier Proteins chemistry, Actins metabolism, Actins genetics, Molecular Dynamics Simulation
- Abstract
Fascin, one of actin bundling proteins, plays an important role in the cross-linking of actin filaments (F-actin). Phosphorylation of Fascin is an important posttranslational modification to affect its structure and function. For example, a phosphomimetic mutation of Fascin-S39D decrease its bundling ability with F-actin significantly. In this paper, we studied the actin-bundling activity of Fascin by using molecular dynamics (MD) simulations and biochemical methods. All single-site mutations from serine/threonine to aspartic acid were mimicked by MD simulations. For five mutants (S146D, S156D, S218D, T239D and S259D), the mutated residues in domain 2 of Fascin were found to form salt-bridge interactions with an adjacent residue, indicating that mutations of these residues could potentially reduce actin-bundling activity. Further, F-actin-bundling assays and immunofluorescence technique showed S146D and T239D to have a strong effect on Fascin bundling with F-actin. Finally, we show that single-site mutations do not change the general shape of Fascin, but local structures near the mutated residues in Fascin-S146D and T239D become unstable, thereby affecting the ability of Fascin to bind with F-actin. These findings suggest that targeting domain 2 of Fascin would be very useful for the drug design. In addition, our study indicates that MD simulation is a useful method to screening which residues on Fascin are important., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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