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Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity

L. pneumophila, an important facultative intracellular bacterium, infects the human lung and environmental protozoa. At least fifteen phospholipases A (PLA) are encoded in its genome. Three of which, namely PlaA, PlaC, and PlaD, belong to the GDSL lipase family abundant in bacteria and higher plants...

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Autores principales: Lang, Christina, Hiller, Miriam, Flieger, Antje
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5701174/
https://www.ncbi.nlm.nih.gov/pubmed/29176577
http://dx.doi.org/10.1038/s41598-017-12796-4
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author Lang, Christina
Hiller, Miriam
Flieger, Antje
author_facet Lang, Christina
Hiller, Miriam
Flieger, Antje
author_sort Lang, Christina
collection PubMed
description L. pneumophila, an important facultative intracellular bacterium, infects the human lung and environmental protozoa. At least fifteen phospholipases A (PLA) are encoded in its genome. Three of which, namely PlaA, PlaC, and PlaD, belong to the GDSL lipase family abundant in bacteria and higher plants. PlaA is a lysophospholipase A (LPLA) that destabilizes the phagosomal membrane in absence of a protective factor. PlaC shows PLA and glycerophospholipid: cholesterol acyltransferase (GCAT) activities which are activated by zinc metalloproteinase ProA via cleavage of a disulphide loop. In this work, we compared GDSL enzyme activities, their secretion, and activation of PlaA. We found that PlaA majorly contributed to LPLA, PlaC to PLA, and both substrate-dependently to GCAT activity. Western blotting revealed that PlaA and PlaC are type II-secreted and both processed by ProA. Interestingly, ProA steeply increased LPLA but diminished GCAT activity of PlaA. Deletion of 20 amino acids within a predicted disulfide loop of PlaA had the same effect. In summary, we propose a model by which ProA processes PlaA via disulfide loop cleavage leading to a steep increase in LPLA activity. Our results help to further characterize the L. pneumophila GDSL hydrolases, particularly PlaA, an enzyme acting in the Legionella-containing phagosome.
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spelling pubmed-57011742017-11-30 Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity Lang, Christina Hiller, Miriam Flieger, Antje Sci Rep Article L. pneumophila, an important facultative intracellular bacterium, infects the human lung and environmental protozoa. At least fifteen phospholipases A (PLA) are encoded in its genome. Three of which, namely PlaA, PlaC, and PlaD, belong to the GDSL lipase family abundant in bacteria and higher plants. PlaA is a lysophospholipase A (LPLA) that destabilizes the phagosomal membrane in absence of a protective factor. PlaC shows PLA and glycerophospholipid: cholesterol acyltransferase (GCAT) activities which are activated by zinc metalloproteinase ProA via cleavage of a disulphide loop. In this work, we compared GDSL enzyme activities, their secretion, and activation of PlaA. We found that PlaA majorly contributed to LPLA, PlaC to PLA, and both substrate-dependently to GCAT activity. Western blotting revealed that PlaA and PlaC are type II-secreted and both processed by ProA. Interestingly, ProA steeply increased LPLA but diminished GCAT activity of PlaA. Deletion of 20 amino acids within a predicted disulfide loop of PlaA had the same effect. In summary, we propose a model by which ProA processes PlaA via disulfide loop cleavage leading to a steep increase in LPLA activity. Our results help to further characterize the L. pneumophila GDSL hydrolases, particularly PlaA, an enzyme acting in the Legionella-containing phagosome. Nature Publishing Group UK 2017-11-24 /pmc/articles/PMC5701174/ /pubmed/29176577 http://dx.doi.org/10.1038/s41598-017-12796-4 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Lang, Christina
Hiller, Miriam
Flieger, Antje
Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title_full Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title_fullStr Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title_full_unstemmed Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title_short Disulfide loop cleavage of Legionella pneumophila PlaA boosts lysophospholipase A activity
title_sort disulfide loop cleavage of legionella pneumophila plaa boosts lysophospholipase a activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5701174/
https://www.ncbi.nlm.nih.gov/pubmed/29176577
http://dx.doi.org/10.1038/s41598-017-12796-4
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