Enhanced production of the mical redox domain for enzymology and F‐actin disassembly assays

Jimok Yoon, Heng Wu, Ruei Jiun Hung, Jonathan R. Terman

Research output: Contribution to journalArticlepeer-review

Abstract

To change their behaviors, cells require actin proteins to assemble together into long pol-ymers/filaments—and so a critical goal is to understand the factors that control this actin filament (F‐actin) assembly and stability. We have identified a family of unusual actin regulators, the MI‐ CALs, which are flavoprotein monooxygenase/hydroxylase enzymes that associate with flavin ad-enine dinucleotide (FAD) and use the co‐enzyme nicotinamide adenine dinucleotide phosphate (NADPH) in Redox reactions. F‐actin is a specific substrate for these MICAL Redox enzymes, which oxidize specific amino acids within actin to destabilize actin filaments. Furthermore, this MICAL-catalyzed reaction is reversed by another family of Redox enzymes (SelR/MsrB enzymes)—thereby revealing a reversible Redox signaling process and biochemical mechanism regulating actin dynam-ics. Interestingly, in addition to the MICALs’ Redox enzymatic portion through which MICALs co-valently modify and affect actin, MICALs have multiple other domains. Less is known about the roles of these other MICAL domains. Here we provide approaches for obtaining high levels of re-combinant protein for the Redox only portion of Mical and demonstrate its catalytic and F‐actin disassembly activity. These results provide a ground state for future work aimed at defining the role of the other domains of Mical — including characterizing their effects on Mical’s Redox enzy-matic and F‐actin disassembly activity.

Original languageEnglish (US)
Article number1991
Pages (from-to)1-17
Number of pages17
JournalInternational journal of molecular sciences
Volume22
Issue number4
DOIs
StatePublished - Feb 2 2021

Keywords

  • Drosophila
  • F‐actin disassembly
  • MICAL1
  • MICAL2
  • MICAL3
  • Oxidoreductase
  • Plexin
  • Repellent
  • Repulsion
  • Semaphorin

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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