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Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays
To change their behaviors, cells require actin proteins to assemble together into long polymers/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 MICALs, which...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922515/ https://www.ncbi.nlm.nih.gov/pubmed/33671465 http://dx.doi.org/10.3390/ijms22041991 |
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author | Yoon, Jimok Wu, Heng Hung, Ruei-Jiun Terman, Jonathan R. |
author_facet | Yoon, Jimok Wu, Heng Hung, Ruei-Jiun Terman, Jonathan R. |
author_sort | Yoon, Jimok |
collection | PubMed |
description | To change their behaviors, cells require actin proteins to assemble together into long polymers/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 MICALs, which are flavoprotein monooxygenase/hydroxylase enzymes that associate with flavin adenine 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 dynamics. Interestingly, in addition to the MICALs’ Redox enzymatic portion through which MICALs covalently 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 recombinant 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 enzymatic and F-actin disassembly activity. |
format | Online Article Text |
id | pubmed-7922515 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-79225152021-03-03 Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays Yoon, Jimok Wu, Heng Hung, Ruei-Jiun Terman, Jonathan R. Int J Mol Sci Article To change their behaviors, cells require actin proteins to assemble together into long polymers/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 MICALs, which are flavoprotein monooxygenase/hydroxylase enzymes that associate with flavin adenine 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 dynamics. Interestingly, in addition to the MICALs’ Redox enzymatic portion through which MICALs covalently 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 recombinant 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 enzymatic and F-actin disassembly activity. MDPI 2021-02-17 /pmc/articles/PMC7922515/ /pubmed/33671465 http://dx.doi.org/10.3390/ijms22041991 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Yoon, Jimok Wu, Heng Hung, Ruei-Jiun Terman, Jonathan R. Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title | Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title_full | Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title_fullStr | Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title_full_unstemmed | Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title_short | Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays |
title_sort | enhanced production of the mical redox domain for enzymology and f-actin disassembly assays |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922515/ https://www.ncbi.nlm.nih.gov/pubmed/33671465 http://dx.doi.org/10.3390/ijms22041991 |
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