Abstract
The MICALs are a family of phylogenetically conserved cytoplasmic proteins that modulate numerous cellular behaviors and play critical roles in semaphorin-plexin signaling. Our recent results have revealed that the MICALs are an unusual family of actin regulatory proteins that use actin filaments (F-actin) as a direct substrate-controlling F-actin dynamics via stereospecific oxidation of conserved methionine (Met44 and Met47) residues within actin. In particular, the MICALs have a highly conserved flavoprotein monooxygenase (redox) enzymatic domain in their N-terminus that directly oxidizes and destabilizes F-actin. Here, we describe methods to characterize MICAL-mediated F-actin disassembly using in vitro assays with purified proteins.
Original language | English (US) |
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Pages (from-to) | 119-128 |
Number of pages | 10 |
Journal | Methods in Molecular Biology |
Volume | 1493 |
DOIs | |
State | Published - 2017 |
Keywords
- Actin sedimentation
- F-actin disassembly
- MICALs
- Oxidoreductase
- Plexin
- Pyrene-actin
ASJC Scopus subject areas
- Molecular Biology
- Genetics