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Structural basis for distinct operational modes and protease activation in AAA+ protease Lon

Substrate-bound structures of AAA+ protein translocases reveal a conserved asymmetric spiral staircase architecture wherein a sequential ATP hydrolysis cycle drives hand-over-hand substrate translocation. However, this configuration is unlikely to represent the full conformational landscape of these...

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Autores principales: Shin, Mia, Puchades, Cristina, Asmita, Ananya, Puri, Neha, Adjei, Eric, Wiseman, R. Luke, Karzai, A. Wali, Lander, Gabriel C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7239648/
https://www.ncbi.nlm.nih.gov/pubmed/32490208
http://dx.doi.org/10.1126/sciadv.aba8404
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author Shin, Mia
Puchades, Cristina
Asmita, Ananya
Puri, Neha
Adjei, Eric
Wiseman, R. Luke
Karzai, A. Wali
Lander, Gabriel C.
author_facet Shin, Mia
Puchades, Cristina
Asmita, Ananya
Puri, Neha
Adjei, Eric
Wiseman, R. Luke
Karzai, A. Wali
Lander, Gabriel C.
author_sort Shin, Mia
collection PubMed
description Substrate-bound structures of AAA+ protein translocases reveal a conserved asymmetric spiral staircase architecture wherein a sequential ATP hydrolysis cycle drives hand-over-hand substrate translocation. However, this configuration is unlikely to represent the full conformational landscape of these enzymes, as biochemical studies suggest distinct conformational states depending on the presence or absence of substrate. Here, we used cryo–electron microscopy to determine structures of the Yersinia pestis Lon AAA+ protease in the absence and presence of substrate, uncovering the mechanistic basis for two distinct operational modes. In the absence of substrate, Lon adopts a left-handed, “open” spiral organization with autoinhibited proteolytic active sites. Upon the addition of substrate, Lon undergoes a reorganization to assemble an enzymatically active, right-handed “closed” conformer with active protease sites. These findings define the mechanistic principles underlying the operational plasticity required for processing diverse protein substrates.
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spelling pubmed-72396482020-06-01 Structural basis for distinct operational modes and protease activation in AAA+ protease Lon Shin, Mia Puchades, Cristina Asmita, Ananya Puri, Neha Adjei, Eric Wiseman, R. Luke Karzai, A. Wali Lander, Gabriel C. Sci Adv Research Articles Substrate-bound structures of AAA+ protein translocases reveal a conserved asymmetric spiral staircase architecture wherein a sequential ATP hydrolysis cycle drives hand-over-hand substrate translocation. However, this configuration is unlikely to represent the full conformational landscape of these enzymes, as biochemical studies suggest distinct conformational states depending on the presence or absence of substrate. Here, we used cryo–electron microscopy to determine structures of the Yersinia pestis Lon AAA+ protease in the absence and presence of substrate, uncovering the mechanistic basis for two distinct operational modes. In the absence of substrate, Lon adopts a left-handed, “open” spiral organization with autoinhibited proteolytic active sites. Upon the addition of substrate, Lon undergoes a reorganization to assemble an enzymatically active, right-handed “closed” conformer with active protease sites. These findings define the mechanistic principles underlying the operational plasticity required for processing diverse protein substrates. American Association for the Advancement of Science 2020-05-20 /pmc/articles/PMC7239648/ /pubmed/32490208 http://dx.doi.org/10.1126/sciadv.aba8404 Text en Copyright © 2020 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). http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (http://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 Research Articles
Shin, Mia
Puchades, Cristina
Asmita, Ananya
Puri, Neha
Adjei, Eric
Wiseman, R. Luke
Karzai, A. Wali
Lander, Gabriel C.
Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title_full Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title_fullStr Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title_full_unstemmed Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title_short Structural basis for distinct operational modes and protease activation in AAA+ protease Lon
title_sort structural basis for distinct operational modes and protease activation in aaa+ protease lon
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7239648/
https://www.ncbi.nlm.nih.gov/pubmed/32490208
http://dx.doi.org/10.1126/sciadv.aba8404
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