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Development of new tools to study membrane-anchored mammalian Atg8 proteins
Mammals conserve multiple mammalian Atg8-family proteins (mATG8s) consisting of GABARAP (GABA type A receptor-associated protein) and MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) subfamilies that tightly bind to autophagic membranes in a membrane-anchored form. These proteins are cru...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240976/ https://www.ncbi.nlm.nih.gov/pubmed/36250672 http://dx.doi.org/10.1080/15548627.2022.2132040 |
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author | Park, Sang-Won Jeon, Pureum Yamasaki, Akinori Lee, Hye Eun Choi, Haneul Mun, Ji Young Jun, Yong-Woo Park, Ju-Hui Lee, Seung-Hwan Lee, Soo-Kyeong Lee, You-Kyung Song, Hyun Kyu Lazarou, Michael Cho, Dong-Hyong Komatsu, Masaaki Noda, Nobuo N. Jang, Deok-Jin Lee, Jin-A |
author_facet | Park, Sang-Won Jeon, Pureum Yamasaki, Akinori Lee, Hye Eun Choi, Haneul Mun, Ji Young Jun, Yong-Woo Park, Ju-Hui Lee, Seung-Hwan Lee, Soo-Kyeong Lee, You-Kyung Song, Hyun Kyu Lazarou, Michael Cho, Dong-Hyong Komatsu, Masaaki Noda, Nobuo N. Jang, Deok-Jin Lee, Jin-A |
author_sort | Park, Sang-Won |
collection | PubMed |
description | Mammals conserve multiple mammalian Atg8-family proteins (mATG8s) consisting of GABARAP (GABA type A receptor-associated protein) and MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) subfamilies that tightly bind to autophagic membranes in a membrane-anchored form. These proteins are crucial for selective autophagy and recruit proteins bearing LC3-interacting region (LIR) motifs. However, because limited research tools are available, information on the specific roles of each membrane-anchored mATG8 in selective autophagy is scarce. In this study, we identified LIR motifs specific to the membrane-anchored form of each mATG8 and characterized the residues critical for their selective interaction using cell-based assays and structural analyses. We then used these selective LIR motifs to develop probes and irreversible deconjugases that targeted selective membrane-anchored mATG8s in the autophagic membrane, revealing that membrane-anchored GABARAP subfamily proteins regulate the aggrephagy of amyotrophic lateral sclerosis-linked protein aggregates. Our tools will be useful for elucidating the functional significance of each mATG8 protein on autophagic membranes in autophagy research. ABBREVIATIONS: A:C autophagic membrane:cytosol; ALS amyotrophic lateral sclerosis; ATG4 autophagy related 4; Atg8 autophagy related 8; BafA1 bafilomycin A(1); BNIP3L/Nix BCL2 interacting protein 3 like; CALCOCO2/NDP52 calcium binding and coiled-coil domain 2; EBSS Earle’s balanced salt solution; GABARAP GABA type A receptor-associated protein; GST glutathione S transferase; HKO hexa knockout; K(d) dissociation constant; LIR LC3-interacting region; MAP1LC3/LC3 microtubule associated protein 1 light chain 3; NLS nuclear localization signal/sequence; PE phosphatidylethanolamine; SpHfl1 Schizosaccharomyces pombeorganic solute transmembrane transporter; SQSTM1/p62 SQSTM1/p62; TARDBP/TDP-43 TAR DNA binding protein; TKO triple knockout |
format | Online Article Text |
id | pubmed-10240976 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-102409762023-06-06 Development of new tools to study membrane-anchored mammalian Atg8 proteins Park, Sang-Won Jeon, Pureum Yamasaki, Akinori Lee, Hye Eun Choi, Haneul Mun, Ji Young Jun, Yong-Woo Park, Ju-Hui Lee, Seung-Hwan Lee, Soo-Kyeong Lee, You-Kyung Song, Hyun Kyu Lazarou, Michael Cho, Dong-Hyong Komatsu, Masaaki Noda, Nobuo N. Jang, Deok-Jin Lee, Jin-A Autophagy Research Paper Mammals conserve multiple mammalian Atg8-family proteins (mATG8s) consisting of GABARAP (GABA type A receptor-associated protein) and MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) subfamilies that tightly bind to autophagic membranes in a membrane-anchored form. These proteins are crucial for selective autophagy and recruit proteins bearing LC3-interacting region (LIR) motifs. However, because limited research tools are available, information on the specific roles of each membrane-anchored mATG8 in selective autophagy is scarce. In this study, we identified LIR motifs specific to the membrane-anchored form of each mATG8 and characterized the residues critical for their selective interaction using cell-based assays and structural analyses. We then used these selective LIR motifs to develop probes and irreversible deconjugases that targeted selective membrane-anchored mATG8s in the autophagic membrane, revealing that membrane-anchored GABARAP subfamily proteins regulate the aggrephagy of amyotrophic lateral sclerosis-linked protein aggregates. Our tools will be useful for elucidating the functional significance of each mATG8 protein on autophagic membranes in autophagy research. ABBREVIATIONS: A:C autophagic membrane:cytosol; ALS amyotrophic lateral sclerosis; ATG4 autophagy related 4; Atg8 autophagy related 8; BafA1 bafilomycin A(1); BNIP3L/Nix BCL2 interacting protein 3 like; CALCOCO2/NDP52 calcium binding and coiled-coil domain 2; EBSS Earle’s balanced salt solution; GABARAP GABA type A receptor-associated protein; GST glutathione S transferase; HKO hexa knockout; K(d) dissociation constant; LIR LC3-interacting region; MAP1LC3/LC3 microtubule associated protein 1 light chain 3; NLS nuclear localization signal/sequence; PE phosphatidylethanolamine; SpHfl1 Schizosaccharomyces pombeorganic solute transmembrane transporter; SQSTM1/p62 SQSTM1/p62; TARDBP/TDP-43 TAR DNA binding protein; TKO triple knockout Taylor & Francis 2022-10-17 /pmc/articles/PMC10240976/ /pubmed/36250672 http://dx.doi.org/10.1080/15548627.2022.2132040 Text en © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Paper Park, Sang-Won Jeon, Pureum Yamasaki, Akinori Lee, Hye Eun Choi, Haneul Mun, Ji Young Jun, Yong-Woo Park, Ju-Hui Lee, Seung-Hwan Lee, Soo-Kyeong Lee, You-Kyung Song, Hyun Kyu Lazarou, Michael Cho, Dong-Hyong Komatsu, Masaaki Noda, Nobuo N. Jang, Deok-Jin Lee, Jin-A Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title | Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title_full | Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title_fullStr | Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title_full_unstemmed | Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title_short | Development of new tools to study membrane-anchored mammalian Atg8 proteins |
title_sort | development of new tools to study membrane-anchored mammalian atg8 proteins |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240976/ https://www.ncbi.nlm.nih.gov/pubmed/36250672 http://dx.doi.org/10.1080/15548627.2022.2132040 |
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