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dc.contributor.authorVasil, Michael L.
dc.contributor.authorStonehouse, Martin J.
dc.contributor.authorVasil, Adriana I.
dc.contributor.authorWadsworth, Sandra J.
dc.contributor.authorGoldfine, Howard
dc.contributor.authorBolcome, Robert E.
dc.contributor.authorChan, Joanne
dc.date.accessioned2011-03-27T20:49:59Z
dc.date.issued2009
dc.identifier.citationVasil, Michael L., Martin J. Stonehouse, Adriana I. Vasil, Sandra J. Wadsworth, Howard Goldfine, Robert E. Bolcome, and Joanne Chan. 2009. A complex extracellular sphingomyelinase of inhibits angiogenesis by selective cytotoxicity to endothelial cells. PLoS Pathogens 5(5): e1000420.en_US
dc.identifier.issn1553-7366en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:4774199
dc.description.abstractThe hemolytic phospholipase C (PlcHR) expressed by Pseudomonas aeruginosa is the original member of a Phosphoesterase Superfamily, which includes phosphorylcholine-specific phospholipases C (PC-PLC) produced by frank and opportunistic pathogens. PlcHR, but not all its family members, is also a potent sphingomyelinase (SMase). Data presented herein indicate that picomolar (pM) concentrations of PlcHR are selectively lethal to endothelial cells (EC). An RGD motif of PlcHR contributes to this selectivity. Peptides containing an RGD motif (i.e., GRGDS), but not control peptides (i.e., GDGRS), block the effects of PlcHR on calcium signaling and cytotoxicity to EC. Moreover, RGD variants of PlcHR (e.g., RGE, KGD) are significantly reduced in their binding and toxicity, but retain the enzymatic activity of the wild type PlcHR. PlcHR also inhibits several EC-dependent in vitro assays (i.e., EC migration, EC invasion, and EC tubule formation), which represent key processes involved in angiogenesis (i.e., formation of new blood vessels from existing vasculature). Finally, the impact of PlcHR in an in vivo model of angiogenesis in transgenic zebrafish, and ones treated with an antisense morpholino to knock down a key blood cell regulator, were evaluated because in vitro assays cannot fully represent the complex processes of angiogenesis. As little as 2 ng/embryo of PlcHR was lethal to ∼50% of EGFP-labeled EC at 6 h after injection of embryos at 48 hpf (hours post-fertilization). An active site mutant of PlcHR (Thr178Ala) exhibited 120-fold reduced inhibitory activity in the EC invasion assay, and 20 ng/embryo elicited no detectable inhibitory activity in the zebrafish model. Taken together, these observations are pertinent to the distinctive vasculitis and poor wound healing associated with P. aeruginosa sepsis and suggest that the potent antiangiogenic properties of PlcHR are worthy of further investigation for the treatment of diseases where angiogenesis contributes pathological conditions (e.g., vascularization of tumors, diabetic retinopathy).en_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi:10.1371/journal.ppat.1000420en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2673038/pdf/en_US
dash.licenseLAA
dc.subjectbiochemistryen_US
dc.subjectprotein chemistryen_US
dc.subjectcell biologyen_US
dc.subjectcell signalingen_US
dc.subjectgenetics and genomicsen_US
dc.subjectdisease modelsen_US
dc.subjectinfectious diseasesen_US
dc.subjectbacterial infectionsen_US
dc.subjectnosocomial and healthcare-associated infectionsen_US
dc.subjectmicrobiologyen_US
dc.subjectcellular microbiology and pathogenesisen_US
dc.titleA Complex Extracellular Sphingomyelinase of Pseudomonas aeruginosa Inhibits Angiogenesis by Selective Cytotoxicity to Endothelial Cellsen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS Pathogensen_US
dash.depositing.authorChan, Joanne
dc.date.available2011-03-27T20:49:59Z
dash.affiliation.otherHMS^Surgery-Children's Hospitalen_US
dc.identifier.doi10.1371/journal.ppat.1000420*
dash.authorsorderedfalse
dash.contributor.affiliatedChan, Joanne


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