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Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
Gentlyase is a bacterial extracellular metalloprotease that is widely applied in cell culture and for tissue dissociation and that belongs to the family of thermolysin-like proteases. The structure of thermolysin has been known since 1972 and that of Bacillus cereus neutral protease since 1992. Howe...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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International Union of Crystallography
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3532130/ https://www.ncbi.nlm.nih.gov/pubmed/23275160 http://dx.doi.org/10.1107/S0907444912041169 |
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author | Ruf, Armin Stihle, Martine Benz, Jörg Schmidt, Manfred Sobek, Harald |
author_facet | Ruf, Armin Stihle, Martine Benz, Jörg Schmidt, Manfred Sobek, Harald |
author_sort | Ruf, Armin |
collection | PubMed |
description | Gentlyase is a bacterial extracellular metalloprotease that is widely applied in cell culture and for tissue dissociation and that belongs to the family of thermolysin-like proteases. The structure of thermolysin has been known since 1972 and that of Bacillus cereus neutral protease since 1992. However, the structure determination of other Bacillus neutral proteases has been hindered by their tendency to cannibalistic autolysis. High calcium conditions that allow the concentration and crystallization of the active Gentlyase metalloprotease without autoproteolysis were identified using thermal fluorescent shift assays. X-ray structures of the protease were solved in the absence and in the presence of the inhibitor phosphoramidon at 1.59 and 1.76 Å resolution, respectively. No domain movement was observed upon inhibitor binding, although such movement is thought to be a general feature of the thermolysin-like protease family. Further analysis of the structure shows that the observed calcium dependency of Gentlyase stability may arise from a partly degenerated calcium site Ca1–2 and a deletion near site Ca3. |
format | Online Article Text |
id | pubmed-3532130 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-35321302013-01-02 Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa Ruf, Armin Stihle, Martine Benz, Jörg Schmidt, Manfred Sobek, Harald Acta Crystallogr D Biol Crystallogr Research Papers Gentlyase is a bacterial extracellular metalloprotease that is widely applied in cell culture and for tissue dissociation and that belongs to the family of thermolysin-like proteases. The structure of thermolysin has been known since 1972 and that of Bacillus cereus neutral protease since 1992. However, the structure determination of other Bacillus neutral proteases has been hindered by their tendency to cannibalistic autolysis. High calcium conditions that allow the concentration and crystallization of the active Gentlyase metalloprotease without autoproteolysis were identified using thermal fluorescent shift assays. X-ray structures of the protease were solved in the absence and in the presence of the inhibitor phosphoramidon at 1.59 and 1.76 Å resolution, respectively. No domain movement was observed upon inhibitor binding, although such movement is thought to be a general feature of the thermolysin-like protease family. Further analysis of the structure shows that the observed calcium dependency of Gentlyase stability may arise from a partly degenerated calcium site Ca1–2 and a deletion near site Ca3. International Union of Crystallography 2013-01-01 2012-12-20 /pmc/articles/PMC3532130/ /pubmed/23275160 http://dx.doi.org/10.1107/S0907444912041169 Text en © Ruf et al. 2013 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Ruf, Armin Stihle, Martine Benz, Jörg Schmidt, Manfred Sobek, Harald Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa |
title | Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
|
title_full | Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
|
title_fullStr | Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
|
title_full_unstemmed | Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
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title_short | Structure of Gentlyase, the neutral metalloprotease of Paenibacillus polymyxa
|
title_sort | structure of gentlyase, the neutral metalloprotease of paenibacillus polymyxa |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3532130/ https://www.ncbi.nlm.nih.gov/pubmed/23275160 http://dx.doi.org/10.1107/S0907444912041169 |
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