Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria

Abstract Sphingolipids, including ceramides, are a diverse group of structurally related lipids composed of a sphingoid base backbone coupled to a fatty acid side chain and modified terminal hydroxyl group. Recently, it has been shown that sphingolipids show antimicrobial activity against a broad ra...

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Autores principales:	Simon Peters, Lena Kaiser, Julian Fink, Fabian Schumacher, Veronika Perschin, Jan Schlegel, Markus Sauer, Christian Stigloher, Burkhard Kleuser, Jürgen Seibel, Alexandra Schubert-Unkmeir
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c1934010a96e45aa82d122a97033f33c
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spelling	oai:doaj.org-article:c1934010a96e45aa82d122a97033f33c2021-12-02T16:23:22ZClick-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria10.1038/s41598-021-83813-w2045-2322https://doaj.org/article/c1934010a96e45aa82d122a97033f33c2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83813-whttps://doaj.org/toc/2045-2322Abstract Sphingolipids, including ceramides, are a diverse group of structurally related lipids composed of a sphingoid base backbone coupled to a fatty acid side chain and modified terminal hydroxyl group. Recently, it has been shown that sphingolipids show antimicrobial activity against a broad range of pathogenic microorganisms. The antimicrobial mechanism, however, remains so far elusive. Here, we introduce ‘click-AT-CLEM’, a labeling technique for correlated light and electron microscopy (CLEM) based on the super-resolution array tomography (srAT) approach and bio-orthogonal click chemistry for imaging of azido-tagged sphingolipids to directly visualize their interaction with the model Gram-negative bacterium Neisseria meningitidis at subcellular level. We observed ultrastructural damage of bacteria and disruption of the bacterial outer membrane induced by two azido-modified sphingolipids by scanning electron microscopy and transmission electron microscopy. Click-AT-CLEM imaging and mass spectrometry clearly revealed efficient incorporation of azido-tagged sphingolipids into the outer membrane of Gram-negative bacteria as underlying cause of their antimicrobial activity.Simon PetersLena KaiserJulian FinkFabian SchumacherVeronika PerschinJan SchlegelMarkus SauerChristian StigloherBurkhard KleuserJürgen SeibelAlexandra Schubert-UnkmeirNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Simon Peters Lena Kaiser Julian Fink Fabian Schumacher Veronika Perschin Jan Schlegel Markus Sauer Christian Stigloher Burkhard Kleuser Jürgen Seibel Alexandra Schubert-Unkmeir Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
description	Abstract Sphingolipids, including ceramides, are a diverse group of structurally related lipids composed of a sphingoid base backbone coupled to a fatty acid side chain and modified terminal hydroxyl group. Recently, it has been shown that sphingolipids show antimicrobial activity against a broad range of pathogenic microorganisms. The antimicrobial mechanism, however, remains so far elusive. Here, we introduce ‘click-AT-CLEM’, a labeling technique for correlated light and electron microscopy (CLEM) based on the super-resolution array tomography (srAT) approach and bio-orthogonal click chemistry for imaging of azido-tagged sphingolipids to directly visualize their interaction with the model Gram-negative bacterium Neisseria meningitidis at subcellular level. We observed ultrastructural damage of bacteria and disruption of the bacterial outer membrane induced by two azido-modified sphingolipids by scanning electron microscopy and transmission electron microscopy. Click-AT-CLEM imaging and mass spectrometry clearly revealed efficient incorporation of azido-tagged sphingolipids into the outer membrane of Gram-negative bacteria as underlying cause of their antimicrobial activity.
format	article
author	Simon Peters Lena Kaiser Julian Fink Fabian Schumacher Veronika Perschin Jan Schlegel Markus Sauer Christian Stigloher Burkhard Kleuser Jürgen Seibel Alexandra Schubert-Unkmeir
author_facet	Simon Peters Lena Kaiser Julian Fink Fabian Schumacher Veronika Perschin Jan Schlegel Markus Sauer Christian Stigloher Burkhard Kleuser Jürgen Seibel Alexandra Schubert-Unkmeir
author_sort	Simon Peters
title	Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
title_short	Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
title_full	Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
title_fullStr	Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
title_full_unstemmed	Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria
title_sort	click-correlative light and electron microscopy (click-at-clem) for imaging and tracking azido-functionalized sphingolipids in bacteria
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/c1934010a96e45aa82d122a97033f33c
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Click-correlative light and electron microscopy (click-AT-CLEM) for imaging and tracking azido-functionalized sphingolipids in bacteria

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