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Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP

The ubiquitin–proteasome system (UPS) is responsible for proteasomal degradation, regulating the half-life of the protein. Deubiquitinating enzymes (DUBs) are components of the UPS and inhibit degradation by removing ubiquitins from protein substrates. Herpesvirus-associated ubiquitin-specific prote...

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Autores principales: Choi, Hae-Seul, Pei, Chang-Zhu, Park, Jun-Hyeok, Kim, Soo-Yeon, Song, Seung-Yeon, Shin, Gyeong-Jin, Baek, Kwang-Hyun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352364/
https://www.ncbi.nlm.nih.gov/pubmed/32545446
http://dx.doi.org/10.3390/cancers12061548
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author Choi, Hae-Seul
Pei, Chang-Zhu
Park, Jun-Hyeok
Kim, Soo-Yeon
Song, Seung-Yeon
Shin, Gyeong-Jin
Baek, Kwang-Hyun
author_facet Choi, Hae-Seul
Pei, Chang-Zhu
Park, Jun-Hyeok
Kim, Soo-Yeon
Song, Seung-Yeon
Shin, Gyeong-Jin
Baek, Kwang-Hyun
author_sort Choi, Hae-Seul
collection PubMed
description The ubiquitin–proteasome system (UPS) is responsible for proteasomal degradation, regulating the half-life of the protein. Deubiquitinating enzymes (DUBs) are components of the UPS and inhibit degradation by removing ubiquitins from protein substrates. Herpesvirus-associated ubiquitin-specific protease (HAUSP) is one such deubiquitinating enzyme and has been closely associated with tumor development. In a previous study, we isolated putative HAUSP binding substrates by two-dimensional electrophoresis (2-DE) and identified them by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis. The analysis showed that pyruvate kinase isoenzyme M2 (PKM2) was likely to be one of the substrates for HAUSP. Further study revealed that PKM2 binds to HAUSP, confirming the interaction between these proteins, and that PKM2 possesses the putative HAUSP binding motif, E or P/AXXS. Therefore, we generated mutant forms of PKM2 S57A, S97A, and S346A, and found that S57A had less binding affinity. In a previous study, we demonstrated that PKM2 is regulated by the UPS, and that HAUSP- as a DUB-acted on PKM2, thus siRNA for HAUSP increases PKM2 ubiquitination. Our present study newly highlights the direct interaction between HAUSP and PKM2.
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spelling pubmed-73523642020-07-15 Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP Choi, Hae-Seul Pei, Chang-Zhu Park, Jun-Hyeok Kim, Soo-Yeon Song, Seung-Yeon Shin, Gyeong-Jin Baek, Kwang-Hyun Cancers (Basel) Article The ubiquitin–proteasome system (UPS) is responsible for proteasomal degradation, regulating the half-life of the protein. Deubiquitinating enzymes (DUBs) are components of the UPS and inhibit degradation by removing ubiquitins from protein substrates. Herpesvirus-associated ubiquitin-specific protease (HAUSP) is one such deubiquitinating enzyme and has been closely associated with tumor development. In a previous study, we isolated putative HAUSP binding substrates by two-dimensional electrophoresis (2-DE) and identified them by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis. The analysis showed that pyruvate kinase isoenzyme M2 (PKM2) was likely to be one of the substrates for HAUSP. Further study revealed that PKM2 binds to HAUSP, confirming the interaction between these proteins, and that PKM2 possesses the putative HAUSP binding motif, E or P/AXXS. Therefore, we generated mutant forms of PKM2 S57A, S97A, and S346A, and found that S57A had less binding affinity. In a previous study, we demonstrated that PKM2 is regulated by the UPS, and that HAUSP- as a DUB-acted on PKM2, thus siRNA for HAUSP increases PKM2 ubiquitination. Our present study newly highlights the direct interaction between HAUSP and PKM2. MDPI 2020-06-12 /pmc/articles/PMC7352364/ /pubmed/32545446 http://dx.doi.org/10.3390/cancers12061548 Text en © 2020 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
Choi, Hae-Seul
Pei, Chang-Zhu
Park, Jun-Hyeok
Kim, Soo-Yeon
Song, Seung-Yeon
Shin, Gyeong-Jin
Baek, Kwang-Hyun
Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title_full Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title_fullStr Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title_full_unstemmed Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title_short Protein Stability of Pyruvate Kinase Isozyme M2 Is Mediated by HAUSP
title_sort protein stability of pyruvate kinase isozyme m2 is mediated by hausp
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352364/
https://www.ncbi.nlm.nih.gov/pubmed/32545446
http://dx.doi.org/10.3390/cancers12061548
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