20.500.12556/RUL-144800 Crystal structure of RahU, an aegerolysin protein from the human pathogen Pseudomonas aeruginosa, and its interaction with membrane ceramide phosphorylethanolamine 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. aegerolysins crystal structure of RahU ceramide phosphorylethanolamine biochemistry lipids membrane lipids membrane proteins microbiology proteins sphingolipids structural biology X-ray crystallography aegerolizini kristalna struktura RahU ceramid fosforiletanolamin true false true Angleški jezik Slovenski jezik Članek v reviji 2023-03-13 14:50:22 2023-03-13 14:50:26 2023-03-14 03:29:56 0000-00-00 00:00:00 2021 0 0 12 str. art. 6572 Vol. 11 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 2045-2322 10.1038/s41598-021-85956-2 56737283 RAZ_Kocar_Eva_2021.pdf RAZ_Kocar_Eva_2021.pdf 1 DA516A551D5EB88D0D52C49AA35880A4 f441bff4d3f63b9d44939080db926b2f5e3ce49d9c8a6406cace0cb7a66ecf6b f62a973e-c1a5-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=167321 https://www.nature.com/articles/s41598-021-85956-2 1 f6db5a17-c1a5-11ed-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=167322 Biotehniška fakulteta Medicinska fakulteta 0 0 0