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Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase

Protein ageing is often mediated by the formation of succinimide intermediates. These short-lived intermediates derive from asparaginyl deamidation and aspartyl dehydration and are rapidly converted into β-aspartyl or D-aspartyl residues. Here we report the presence of a highly stable succinimide in...

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Autores principales: Kumar, Sanjeev, Prakash, Sunita, Gupta, Kallol, Dongre, Aparna, Balaram, Padmanabhan, Balaram, Hemalatha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5052720/
https://www.ncbi.nlm.nih.gov/pubmed/27677693
http://dx.doi.org/10.1038/ncomms12798
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author Kumar, Sanjeev
Prakash, Sunita
Gupta, Kallol
Dongre, Aparna
Balaram, Padmanabhan
Balaram, Hemalatha
author_facet Kumar, Sanjeev
Prakash, Sunita
Gupta, Kallol
Dongre, Aparna
Balaram, Padmanabhan
Balaram, Hemalatha
author_sort Kumar, Sanjeev
collection PubMed
description Protein ageing is often mediated by the formation of succinimide intermediates. These short-lived intermediates derive from asparaginyl deamidation and aspartyl dehydration and are rapidly converted into β-aspartyl or D-aspartyl residues. Here we report the presence of a highly stable succinimide intermediate in the glutaminase subunit of GMP synthetase from the hyperthermophile Methanocaldoccocus jannaschii. By comparing the biophysical properties of the wild-type protein and of several mutants, we show that the presence of succinimide increases the structural stability of the glutaminase subunit. The protein bearing this modification in fact remains folded at 100 °C and in 8 M guanidinium chloride. Mutation of the residue following the reactive asparagine provides insight into the factors that contribute to the hydrolytic stability of the succinimide. Our findings suggest that sequences that stabilize succinimides from hydrolysis may be evolutionarily selected to confer extreme thermal stability.
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spelling pubmed-50527202016-10-21 Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase Kumar, Sanjeev Prakash, Sunita Gupta, Kallol Dongre, Aparna Balaram, Padmanabhan Balaram, Hemalatha Nat Commun Article Protein ageing is often mediated by the formation of succinimide intermediates. These short-lived intermediates derive from asparaginyl deamidation and aspartyl dehydration and are rapidly converted into β-aspartyl or D-aspartyl residues. Here we report the presence of a highly stable succinimide intermediate in the glutaminase subunit of GMP synthetase from the hyperthermophile Methanocaldoccocus jannaschii. By comparing the biophysical properties of the wild-type protein and of several mutants, we show that the presence of succinimide increases the structural stability of the glutaminase subunit. The protein bearing this modification in fact remains folded at 100 °C and in 8 M guanidinium chloride. Mutation of the residue following the reactive asparagine provides insight into the factors that contribute to the hydrolytic stability of the succinimide. Our findings suggest that sequences that stabilize succinimides from hydrolysis may be evolutionarily selected to confer extreme thermal stability. Nature Publishing Group 2016-09-28 /pmc/articles/PMC5052720/ /pubmed/27677693 http://dx.doi.org/10.1038/ncomms12798 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Kumar, Sanjeev
Prakash, Sunita
Gupta, Kallol
Dongre, Aparna
Balaram, Padmanabhan
Balaram, Hemalatha
Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title_full Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title_fullStr Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title_full_unstemmed Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title_short Unexpected functional implication of a stable succinimide in the structural stability of Methanocaldococcus jannaschii glutaminase
title_sort unexpected functional implication of a stable succinimide in the structural stability of methanocaldococcus jannaschii glutaminase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5052720/
https://www.ncbi.nlm.nih.gov/pubmed/27677693
http://dx.doi.org/10.1038/ncomms12798
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