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Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination

BACKGROUND: Mutations in the parkin gene, which encodes a ubiquitin ligase (E3), are a major cause of autosomal recessive parkinsonism. Although parkin-mediated ubiquitination was initially linked to protein degradation, accumulating evidence suggests that the enzyme is capable of catalyzing multipl...

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Autores principales: Chew, Katherine C. M., Matsuda, Noriyuki, Saisho, Keiko, Lim, Grace G. Y., Chai, Chou, Tan, Hui-Mei, Tanaka, Keiji, Lim, Kah-Leong
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3100294/
https://www.ncbi.nlm.nih.gov/pubmed/21625422
http://dx.doi.org/10.1371/journal.pone.0019720
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author Chew, Katherine C. M.
Matsuda, Noriyuki
Saisho, Keiko
Lim, Grace G. Y.
Chai, Chou
Tan, Hui-Mei
Tanaka, Keiji
Lim, Kah-Leong
author_facet Chew, Katherine C. M.
Matsuda, Noriyuki
Saisho, Keiko
Lim, Grace G. Y.
Chai, Chou
Tan, Hui-Mei
Tanaka, Keiji
Lim, Kah-Leong
author_sort Chew, Katherine C. M.
collection PubMed
description BACKGROUND: Mutations in the parkin gene, which encodes a ubiquitin ligase (E3), are a major cause of autosomal recessive parkinsonism. Although parkin-mediated ubiquitination was initially linked to protein degradation, accumulating evidence suggests that the enzyme is capable of catalyzing multiple forms of ubiquitin modifications including monoubiquitination, K48- and K63-linked polyubiquitination. In this study, we sought to understand how a single enzyme could exhibit such multifunctional catalytic properties. METHODS AND FINDINGS: By means of in vitro ubiquitination assays coupled with mass spectrometry analysis, we were surprised to find that parkin is apparently capable of mediating E2-independent protein ubiquitination in vitro, an unprecedented activity exhibited by an E3 member. Interestingly, whereas full length parkin catalyzes solely monoubiquitination regardless of the presence or absence of E2, a truncated parkin mutant containing only the catalytic moiety supports both E2-independent and E2-dependent assembly of ubiquitin chains. CONCLUSIONS: Our results here suggest a complex regulation of parkin's activity and may help to explain how a single enzyme like parkin could mediate diverse forms of ubiquitination.
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spelling pubmed-31002942011-05-27 Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination Chew, Katherine C. M. Matsuda, Noriyuki Saisho, Keiko Lim, Grace G. Y. Chai, Chou Tan, Hui-Mei Tanaka, Keiji Lim, Kah-Leong PLoS One Research Article BACKGROUND: Mutations in the parkin gene, which encodes a ubiquitin ligase (E3), are a major cause of autosomal recessive parkinsonism. Although parkin-mediated ubiquitination was initially linked to protein degradation, accumulating evidence suggests that the enzyme is capable of catalyzing multiple forms of ubiquitin modifications including monoubiquitination, K48- and K63-linked polyubiquitination. In this study, we sought to understand how a single enzyme could exhibit such multifunctional catalytic properties. METHODS AND FINDINGS: By means of in vitro ubiquitination assays coupled with mass spectrometry analysis, we were surprised to find that parkin is apparently capable of mediating E2-independent protein ubiquitination in vitro, an unprecedented activity exhibited by an E3 member. Interestingly, whereas full length parkin catalyzes solely monoubiquitination regardless of the presence or absence of E2, a truncated parkin mutant containing only the catalytic moiety supports both E2-independent and E2-dependent assembly of ubiquitin chains. CONCLUSIONS: Our results here suggest a complex regulation of parkin's activity and may help to explain how a single enzyme like parkin could mediate diverse forms of ubiquitination. Public Library of Science 2011-05-23 /pmc/articles/PMC3100294/ /pubmed/21625422 http://dx.doi.org/10.1371/journal.pone.0019720 Text en Chew et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chew, Katherine C. M.
Matsuda, Noriyuki
Saisho, Keiko
Lim, Grace G. Y.
Chai, Chou
Tan, Hui-Mei
Tanaka, Keiji
Lim, Kah-Leong
Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title_full Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title_fullStr Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title_full_unstemmed Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title_short Parkin Mediates Apparent E2-Independent Monoubiquitination In Vitro and Contains an Intrinsic Activity That Catalyzes Polyubiquitination
title_sort parkin mediates apparent e2-independent monoubiquitination in vitro and contains an intrinsic activity that catalyzes polyubiquitination
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3100294/
https://www.ncbi.nlm.nih.gov/pubmed/21625422
http://dx.doi.org/10.1371/journal.pone.0019720
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