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Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP
Site-2 proteases are a conserved family of intramembrane proteases that cleave transmembrane substrates to regulate signal transduction and maintain proteostasis. Here, we elucidated crystal structures of inhibitor-bound forms of bacterial site-2 proteases including Escherichia coli RseP. Structure-...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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American Association for the Advancement of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9401612/ https://www.ncbi.nlm.nih.gov/pubmed/36001659 http://dx.doi.org/10.1126/sciadv.abp9011 |
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author | Imaizumi, Yuki Takanuki, Kazunori Miyake, Takuya Takemoto, Mizuki Hirata, Kunio Hirose, Mika Oi, Rika Kobayashi, Tatsuya Miyoshi, Kenichi Aruga, Rie Yokoyama, Tatsuhiko Katagiri, Shizuka Matsuura, Hiroaki Iwasaki, Kenji Kato, Takayuki Kaneko, Mika K. Kato, Yukinari Tajiri, Michiko Akashi, Satoko Nureki, Osamu Hizukuri, Yohei Akiyama, Yoshinori Nogi, Terukazu |
author_facet | Imaizumi, Yuki Takanuki, Kazunori Miyake, Takuya Takemoto, Mizuki Hirata, Kunio Hirose, Mika Oi, Rika Kobayashi, Tatsuya Miyoshi, Kenichi Aruga, Rie Yokoyama, Tatsuhiko Katagiri, Shizuka Matsuura, Hiroaki Iwasaki, Kenji Kato, Takayuki Kaneko, Mika K. Kato, Yukinari Tajiri, Michiko Akashi, Satoko Nureki, Osamu Hizukuri, Yohei Akiyama, Yoshinori Nogi, Terukazu |
author_sort | Imaizumi, Yuki |
collection | PubMed |
description | Site-2 proteases are a conserved family of intramembrane proteases that cleave transmembrane substrates to regulate signal transduction and maintain proteostasis. Here, we elucidated crystal structures of inhibitor-bound forms of bacterial site-2 proteases including Escherichia coli RseP. Structure-based chemical modification and cross-linking experiments indicated that the RseP domains surrounding the active center undergo conformational changes to expose the substrate-binding site, suggesting that RseP has a gating mechanism to regulate substrate entry. Furthermore, mutational analysis suggests that a conserved electrostatic linkage between the transmembrane and peripheral membrane-associated domains mediates the conformational changes. In vivo cleavage assays also support that the substrate transmembrane helix is unwound by strand addition to the intramembrane β sheet of RseP and is clamped by a conserved asparagine residue at the active center for efficient cleavage. This mechanism underlying the substrate binding, i.e., unwinding and clamping, appears common across distinct families of intramembrane proteases that cleave transmembrane segments. |
format | Online Article Text |
id | pubmed-9401612 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Association for the Advancement of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-94016122022-08-26 Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP Imaizumi, Yuki Takanuki, Kazunori Miyake, Takuya Takemoto, Mizuki Hirata, Kunio Hirose, Mika Oi, Rika Kobayashi, Tatsuya Miyoshi, Kenichi Aruga, Rie Yokoyama, Tatsuhiko Katagiri, Shizuka Matsuura, Hiroaki Iwasaki, Kenji Kato, Takayuki Kaneko, Mika K. Kato, Yukinari Tajiri, Michiko Akashi, Satoko Nureki, Osamu Hizukuri, Yohei Akiyama, Yoshinori Nogi, Terukazu Sci Adv Biomedicine and Life Sciences Site-2 proteases are a conserved family of intramembrane proteases that cleave transmembrane substrates to regulate signal transduction and maintain proteostasis. Here, we elucidated crystal structures of inhibitor-bound forms of bacterial site-2 proteases including Escherichia coli RseP. Structure-based chemical modification and cross-linking experiments indicated that the RseP domains surrounding the active center undergo conformational changes to expose the substrate-binding site, suggesting that RseP has a gating mechanism to regulate substrate entry. Furthermore, mutational analysis suggests that a conserved electrostatic linkage between the transmembrane and peripheral membrane-associated domains mediates the conformational changes. In vivo cleavage assays also support that the substrate transmembrane helix is unwound by strand addition to the intramembrane β sheet of RseP and is clamped by a conserved asparagine residue at the active center for efficient cleavage. This mechanism underlying the substrate binding, i.e., unwinding and clamping, appears common across distinct families of intramembrane proteases that cleave transmembrane segments. American Association for the Advancement of Science 2022-08-24 /pmc/articles/PMC9401612/ /pubmed/36001659 http://dx.doi.org/10.1126/sciadv.abp9011 Text en Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). https://creativecommons.org/licenses/by-nc/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (https://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited. |
spellingShingle | Biomedicine and Life Sciences Imaizumi, Yuki Takanuki, Kazunori Miyake, Takuya Takemoto, Mizuki Hirata, Kunio Hirose, Mika Oi, Rika Kobayashi, Tatsuya Miyoshi, Kenichi Aruga, Rie Yokoyama, Tatsuhiko Katagiri, Shizuka Matsuura, Hiroaki Iwasaki, Kenji Kato, Takayuki Kaneko, Mika K. Kato, Yukinari Tajiri, Michiko Akashi, Satoko Nureki, Osamu Hizukuri, Yohei Akiyama, Yoshinori Nogi, Terukazu Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title | Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title_full | Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title_fullStr | Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title_full_unstemmed | Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title_short | Mechanistic insights into intramembrane proteolysis by E. coli site-2 protease homolog RseP |
title_sort | mechanistic insights into intramembrane proteolysis by e. coli site-2 protease homolog rsep |
topic | Biomedicine and Life Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9401612/ https://www.ncbi.nlm.nih.gov/pubmed/36001659 http://dx.doi.org/10.1126/sciadv.abp9011 |
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