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dc.contributor.authorDiwo, Maurice
dc.contributor.authorMichel, Wiebke
dc.contributor.authorAurass, Philipp
dc.contributor.authorKuhle-Keindorf, Katja
dc.contributor.authorPippel, Jan
dc.contributor.authorKrausze, Joern
dc.contributor.authorWamp, Sabrina
dc.contributor.authorLang, Christina
dc.contributor.authorBlankenfeldt, Wulf
dc.contributor.authorFlieger, Antje
dc.date.accessioned2021-07-20T12:47:27Z
dc.date.available2021-07-20T12:47:27Z
dc.date.issued2021-06-01
dc.identifier.citationProc Natl Acad Sci U S A. 2021 Jun 8;118(23):e2017046118. doi: 10.1073/pnas.2017046118.en_US
dc.identifier.pmid34074754
dc.identifier.doi10.1073/pnas.2017046118
dc.identifier.urihttp://hdl.handle.net/10033/622946
dc.description.abstractThe virulence factor PlaB promotes lung colonization, tissue destruction, and intracellular replication of Legionella pneumophila, the causative agent of Legionnaires' disease. It is a highly active phospholipase exposed at the bacterial surface and shows an extraordinary activation mechanism by tetramer deoligomerization. To unravel the molecular basis for enzyme activation and localization, we determined the crystal structure of PlaB in its tetrameric form. We found that the tetramer is a dimer of identical dimers, and a monomer consists of an N-terminal α/β-hydrolase domain expanded by two noncanonical two-stranded β-sheets, β-6/β-7 and β-9/β-10. The C-terminal domain reveals a fold displaying a bilobed β-sandwich with a hook structure required for dimer formation and structural complementation of the enzymatic domain in the neighboring monomer. This highlights the dimer as the active form. Δβ-9/β-10 mutants showed a decrease in the tetrameric fraction and altered activity profiles. The variant also revealed restricted binding to membranes resulting in mislocalization and bacterial lysis. Unexpectedly, we observed eight NAD(H) molecules at the dimer/dimer interface, suggesting that these molecules stabilize the tetramer and hence lead to enzyme inactivation. Indeed, addition of NAD(H) increased the fraction of the tetramer and concomitantly reduced activity. Together, these data reveal structural elements and an unprecedented NAD(H)-mediated tetramerization mechanism required for spatial and enzymatic control of a phospholipase virulence factor. The allosteric regulatory process identified here is suited to fine tune PlaB in a way that protects Legionella pneumophila from self-inflicted lysis while ensuring its activity at the pathogen-host interface.en_US
dc.language.isoenen_US
dc.publisherNational Academy of Sciencesen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectactivationen_US
dc.subjectdeoligomerizationen_US
dc.subjectmembrane associationen_US
dc.subjectnicotinamide dinucleotideen_US
dc.subjectphospholipaseen_US
dc.titleNAD(H)-mediated tetramerization controls the activity of phospholipase PlaB.en_US
dc.typeArticleen_US
dc.identifier.eissn1091-6490
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalProceedings of the National Academy of Sciences of the United States of Americaen_US
dc.source.volume118
dc.source.issue23
refterms.dateFOA2021-07-20T12:47:28Z
dc.source.journaltitleProceedings of the National Academy of Sciences of the United States of America
dc.source.countryUnited States


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