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A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)

Lipoprotein modification is an essential process in Gram-negative bacteria. The action of three integral membrane proteins that catalyze the transfer of fatty acids derived from membrane phospholipids or cleave the signal peptide of the lipoprotein substrate result in the formation of mature triacyl...

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Autores principales: Nozeret, Karine, Boucharlat, Alix, Agou, Fabrice, Buddelmeijer, Nienke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828757/
https://www.ncbi.nlm.nih.gov/pubmed/31685855
http://dx.doi.org/10.1038/s41598-019-52106-8
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author Nozeret, Karine
Boucharlat, Alix
Agou, Fabrice
Buddelmeijer, Nienke
author_facet Nozeret, Karine
Boucharlat, Alix
Agou, Fabrice
Buddelmeijer, Nienke
author_sort Nozeret, Karine
collection PubMed
description Lipoprotein modification is an essential process in Gram-negative bacteria. The action of three integral membrane proteins that catalyze the transfer of fatty acids derived from membrane phospholipids or cleave the signal peptide of the lipoprotein substrate result in the formation of mature triacylated proteins. Inactivation of the enzymes leads to mis-localization of immature lipoproteins and consequently cell death. Biochemical studies and the development of in vitro assays are challenging due to the fact that the enzymes and substrates are all membrane-embedded proteins difficult to overproduce and purify. Here we describe a sensitive fluorescence-based assay to monitor bacterial apolipoprotein N-acyltransferase activity.
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spelling pubmed-68287572019-11-12 A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt) Nozeret, Karine Boucharlat, Alix Agou, Fabrice Buddelmeijer, Nienke Sci Rep Article Lipoprotein modification is an essential process in Gram-negative bacteria. The action of three integral membrane proteins that catalyze the transfer of fatty acids derived from membrane phospholipids or cleave the signal peptide of the lipoprotein substrate result in the formation of mature triacylated proteins. Inactivation of the enzymes leads to mis-localization of immature lipoproteins and consequently cell death. Biochemical studies and the development of in vitro assays are challenging due to the fact that the enzymes and substrates are all membrane-embedded proteins difficult to overproduce and purify. Here we describe a sensitive fluorescence-based assay to monitor bacterial apolipoprotein N-acyltransferase activity. Nature Publishing Group UK 2019-11-04 /pmc/articles/PMC6828757/ /pubmed/31685855 http://dx.doi.org/10.1038/s41598-019-52106-8 Text en © The Author(s) 2019 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
Nozeret, Karine
Boucharlat, Alix
Agou, Fabrice
Buddelmeijer, Nienke
A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title_full A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title_fullStr A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title_full_unstemmed A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title_short A sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein Apolipoprotein N-acyltransferase (Lnt)
title_sort sensitive fluorescence-based assay to monitor enzymatic activity of the essential integral membrane protein apolipoprotein n-acyltransferase (lnt)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828757/
https://www.ncbi.nlm.nih.gov/pubmed/31685855
http://dx.doi.org/10.1038/s41598-019-52106-8
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