Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine

Abstract Aegerolysins are proteins produced by bacteria, fungi, plants and protozoa. The most studied fungal aegerolysins share a common property of interacting with membranes enriched with cholesterol in combination with either sphingomyelin or ceramide phosphorylethanolamine (CPE), major sphingoli...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Eva Kočar, Tea Lenarčič, Vesna Hodnik, Anastasija Panevska, Yunjie Huang, Gregor Bajc, Rok Kostanjšek, Anjaparavanda P. Naren, Peter Maček, Gregor Anderluh, Kristina Sepčić, Marjetka Podobnik, Matej Butala
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/bf82f07acaef4c448d00464a1303d152
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:bf82f07acaef4c448d00464a1303d152
record_format	dspace
spelling	oai:doaj.org-article:bf82f07acaef4c448d00464a1303d1522021-12-02T16:35:56ZCrystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine10.1038/s41598-021-85956-22045-2322https://doaj.org/article/bf82f07acaef4c448d00464a1303d1522021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85956-2https://doaj.org/toc/2045-2322Abstract Aegerolysins are proteins produced by bacteria, fungi, plants and protozoa. The most studied fungal aegerolysins share a common property of interacting with membranes enriched with cholesterol in combination with either sphingomyelin or ceramide phosphorylethanolamine (CPE), major sphingolipids in the cell membranes of vertebrates and invertebrates, respectively. However, genome analyses show a particularly high frequency of aegerolysin genes in bacteria, including the pathogenic genera Pseudomonas and Vibrio; these are human pathogens of high clinical relevance and can thrive in a variety of other species. The knowledge on bacterial aegerolysin-lipid interactions is scarce. We show that Pseudomonas aeruginosa aegerolysin RahU interacts with CPE, but not with sphingomyelin-enriched artificial membranes, and that RahU interacts with the insect cell line producing CPE. We report crystal structures of RahU alone and in complex with tris(hydroxymethyl)aminomethane (Tris), which, like the phosphorylethanolamine head group of CPE, contains a primary amine. The RahU structures reveal that the two loops proximal to the amino terminus form a cavity that accommodates Tris, and that the flexibility of these two loops is important for this interaction. We show that Tris interferes with CPE-enriched membranes for binding to RahU, implying on the importance of the ligand cavity between the loops and its proximity in RahU membrane interaction. We further support this by studying the interaction of single amino acid substitution mutants of RahU with the CPE-enriched membranes. Our results thus represent a starting point for a better understanding of the role of P. aeruginosa RahU, and possibly other bacterial aegerolysins, in bacterial interactions with other organisms.Eva KočarTea LenarčičVesna HodnikAnastasija PanevskaYunjie HuangGregor BajcRok KostanjšekAnjaparavanda P. NarenPeter MačekGregor AnderluhKristina SepčićMarjetka PodobnikMatej ButalaNature 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 Eva Kočar Tea Lenarčič Vesna Hodnik Anastasija Panevska Yunjie Huang Gregor Bajc Rok Kostanjšek Anjaparavanda P. Naren Peter Maček Gregor Anderluh Kristina Sepčić Marjetka Podobnik Matej Butala Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
description	Abstract Aegerolysins are proteins produced by bacteria, fungi, plants and protozoa. The most studied fungal aegerolysins share a common property of interacting with membranes enriched with cholesterol in combination with either sphingomyelin or ceramide phosphorylethanolamine (CPE), major sphingolipids in the cell membranes of vertebrates and invertebrates, respectively. However, genome analyses show a particularly high frequency of aegerolysin genes in bacteria, including the pathogenic genera Pseudomonas and Vibrio; these are human pathogens of high clinical relevance and can thrive in a variety of other species. The knowledge on bacterial aegerolysin-lipid interactions is scarce. We show that Pseudomonas aeruginosa aegerolysin RahU interacts with CPE, but not with sphingomyelin-enriched artificial membranes, and that RahU interacts with the insect cell line producing CPE. We report crystal structures of RahU alone and in complex with tris(hydroxymethyl)aminomethane (Tris), which, like the phosphorylethanolamine head group of CPE, contains a primary amine. The RahU structures reveal that the two loops proximal to the amino terminus form a cavity that accommodates Tris, and that the flexibility of these two loops is important for this interaction. We show that Tris interferes with CPE-enriched membranes for binding to RahU, implying on the importance of the ligand cavity between the loops and its proximity in RahU membrane interaction. We further support this by studying the interaction of single amino acid substitution mutants of RahU with the CPE-enriched membranes. Our results thus represent a starting point for a better understanding of the role of P. aeruginosa RahU, and possibly other bacterial aegerolysins, in bacterial interactions with other organisms.
format	article
author	Eva Kočar Tea Lenarčič Vesna Hodnik Anastasija Panevska Yunjie Huang Gregor Bajc Rok Kostanjšek Anjaparavanda P. Naren Peter Maček Gregor Anderluh Kristina Sepčić Marjetka Podobnik Matej Butala
author_facet	Eva Kočar Tea Lenarčič Vesna Hodnik Anastasija Panevska Yunjie Huang Gregor Bajc Rok Kostanjšek Anjaparavanda P. Naren Peter Maček Gregor Anderluh Kristina Sepčić Marjetka Podobnik Matej Butala
author_sort	Eva Kočar
title	Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
title_short	Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
title_full	Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
title_fullStr	Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
title_full_unstemmed	Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
title_sort	crystal structure of rahu, an aegerolysin protein from the human pathogen pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/bf82f07acaef4c448d00464a1303d152
work_keys_str_mv	AT evakocar crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT tealenarcic crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT vesnahodnik crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT anastasijapanevska crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT yunjiehuang crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT gregorbajc crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT rokkostanjsek crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT anjaparavandapnaren crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT petermacek crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT gregoranderluh crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT kristinasepcic crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT marjetkapodobnik crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine AT matejbutala crystalstructureofrahuanaegerolysinproteinfromthehumanpathogenpseudomonasaeruginosaanditsinteractionwithmembraneceramidephosphorylethanolamine
_version_	1718383716108075008

Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine

Ejemplares similares