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Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability
The lipase produced by Burkholderia glumae folds spontaneously into an inactive near-native state and requires a periplasmic chaperone to reach its final active and secretion-competent fold. The B. glumae lipase-specific foldase (Lif) is classified as a member of the steric-chaperone family of which...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3352829/ https://www.ncbi.nlm.nih.gov/pubmed/22615867 http://dx.doi.org/10.1371/journal.pone.0036999 |
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author | Pauwels, Kris Sanchez del Pino, Manuel M. Feller, Georges Van Gelder, Patrick |
author_facet | Pauwels, Kris Sanchez del Pino, Manuel M. Feller, Georges Van Gelder, Patrick |
author_sort | Pauwels, Kris |
collection | PubMed |
description | The lipase produced by Burkholderia glumae folds spontaneously into an inactive near-native state and requires a periplasmic chaperone to reach its final active and secretion-competent fold. The B. glumae lipase-specific foldase (Lif) is classified as a member of the steric-chaperone family of which the propeptides of α-lytic protease and subtilisin are the best known representatives. Steric chaperones play a key role in conferring kinetic stability to proteins. However, until present there was no solid experimental evidence that Lif-dependent lipases are kinetically trapped enzymes. By combining thermal denaturation studies with proteolytic resistance experiments and the description of distinct folding intermediates, we demonstrate that the native lipase has a kinetically stable conformation. We show that a newly discovered molten globule-like conformation has distinct properties that clearly differ from those of the near-native intermediate state. The folding fingerprint of Lif-dependent lipases is put in the context of the protease-prodomain system and the comparison reveals clear differences that render the lipase-Lif systems unique. Limited proteolysis unveils structural differences between the near-native intermediate and the native conformation and sets the stage to shed light onto the nature of the kinetic barrier. |
format | Online Article Text |
id | pubmed-3352829 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33528292012-05-21 Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability Pauwels, Kris Sanchez del Pino, Manuel M. Feller, Georges Van Gelder, Patrick PLoS One Research Article The lipase produced by Burkholderia glumae folds spontaneously into an inactive near-native state and requires a periplasmic chaperone to reach its final active and secretion-competent fold. The B. glumae lipase-specific foldase (Lif) is classified as a member of the steric-chaperone family of which the propeptides of α-lytic protease and subtilisin are the best known representatives. Steric chaperones play a key role in conferring kinetic stability to proteins. However, until present there was no solid experimental evidence that Lif-dependent lipases are kinetically trapped enzymes. By combining thermal denaturation studies with proteolytic resistance experiments and the description of distinct folding intermediates, we demonstrate that the native lipase has a kinetically stable conformation. We show that a newly discovered molten globule-like conformation has distinct properties that clearly differ from those of the near-native intermediate state. The folding fingerprint of Lif-dependent lipases is put in the context of the protease-prodomain system and the comparison reveals clear differences that render the lipase-Lif systems unique. Limited proteolysis unveils structural differences between the near-native intermediate and the native conformation and sets the stage to shed light onto the nature of the kinetic barrier. Public Library of Science 2012-05-15 /pmc/articles/PMC3352829/ /pubmed/22615867 http://dx.doi.org/10.1371/journal.pone.0036999 Text en Pauwels 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 Pauwels, Kris Sanchez del Pino, Manuel M. Feller, Georges Van Gelder, Patrick Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title | Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title_full | Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title_fullStr | Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title_full_unstemmed | Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title_short | Decoding the Folding of Burkholderia glumae Lipase: Folding Intermediates En Route to Kinetic Stability |
title_sort | decoding the folding of burkholderia glumae lipase: folding intermediates en route to kinetic stability |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3352829/ https://www.ncbi.nlm.nih.gov/pubmed/22615867 http://dx.doi.org/10.1371/journal.pone.0036999 |
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