title: Ganglioside embedded in reconstituted lipoprotein binds cholera toxin with elevated affinity.
abstract: The ability to exogenously present cell-surface receptors in high-affinity conformations in a synthetic system offers an opportunity to provide host cells with protection from pathogenic toxins. This strategy requires improvement of the synthetic receptor binding affinity against its native counterpart, particularly with polyvalent toxins where clustering of membrane receptors can hinder binding. Here we demonstrate that reconstituted lipoprotein, nanometer-sized discoidal lipid bilayers bounded by apolipoprotein and functionalized by incorporation of pathogen receptors, provides a means to enhance toxin-receptor binding through molecular-level control over the receptor microenvironment (specifically, its rigidity, composition, and heterogeneity). Using a Foerster Resonance Energy Transfer (FRET)-based assay, we found that reconstituted lipoprotein incorporating low concentrations of ganglioside monosialotetrahexosylganglioside (GM1) binds polymeric cholera toxin with significantly higher affinity than liposomes or supported lipid bilayers, most likely a result of the enhanced control over receptor clustering provided by the lipoprotein platform. Using wide-area epifluorescence, we found that this enhanced binding capacity can be effectively utilized to divert cholera toxin away from populations of healthy mammalian cells. In summary, we found that reconstitutions of high-density lipoprotein can be engineered to include specific pathogen receptors; that their pathogen binding affinity is altered, presumably due to attenuation of receptor aggregation; and that these assemblies are effective at protecting cells from biological toxins.
authors.affiliation: Department of Applied Science, University of California, Davis, CA 95616, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Bricarello
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Mills
authors.forename: Jitka
authors.initials: J
authors.lastname: Petrlova
authors.forename: John C
authors.initials: JC
authors.lastname: Voss
authors.forename: Atul N
authors.initials: AN
authors.lastname: Parikh
mesh_terms: D016632:Apolipoprotein A-I / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005732:Gangliosides / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D055213:Retinal Pigment Epithelium / Q000166:cytology
chemical_list: C505693:APOA1 protein, human; D016632:Apolipoprotein A-I; D005732:Gangliosides; D008051:Lipid Bilayers; D008074:Lipoproteins; D008081:Liposomes; D002772:Cholera Toxin
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