title: The interaction of lipodepsipeptide toxins from Pseudomonas syringae pv. syringae with biological and model membranes: a comparison of syringotoxin, syringomycin, and two syringopeptins.
abstract: Pseudomonas syringae pv. syringae produces two groups of cyclic lipodepsipeptides (LDPs): the nona-peptides syringomycins, syringostatins, and syringotoxin (ST), and the more complex syringopeptins composed of either 22 or 25 amino acid residues (SP22 and SP25). Both classes of peptides significantly contribute to bacterial pathogenesis and their primary target of action seems to be the plasma membrane. We studied and compared the activity of some members of these two classes of LDPs on red blood cells and on model membranes (monolayers and unilamellar vesicles). All peptides induced red blood cell hemolysis. The mechanism was apparently that of a colloid-osmotic shock caused by the formation of pores, as it could be prevented by osmoticants of adequate size. Application of the Renkin equation indicated a radius of approximately 1 nm for the lesions formed by syringopeptins SP22A and SP25A, whereas those formed by syringomycin E (SRE) had a variable, dose-dependent size ranging from 0.7 up to 1.7 nm. All tested LDPs displayed surface activity, forming peptide monolayers with average molecular areas of 1.2 nm2 (SRE), 1.5 nm2 (SP22A), and 1.3 nm2 (SP25A). They also partitioned into preformed lipid monolayers occupying molecular areas that ranged from 0.6 to 1.7 nm2 depending on the peptide and the lipid composition of the film. These LDPs formed channels in lipid vesicles as indicated by the release of an entrapped fluorescent dye (calcein). The extent of permeabilization was dependent on the concentration of the peptide and the composition of the lipid vesicles, with a preference for those containing a sterol. From the dose dependence of the permeabilization it was inferred that LDPs increased membrane permeability by forming oligomeric channels containing from four to seven monomers. On average, syringopeptin oligomers were smaller than SRE and ST oligomers.
authors.affiliation: CNR-ITC Centro Fisica Stati Aggregati, Povo, Trento, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Dalla Serra
authors.forename: G
authors.initials: G
authors.lastname: Fagiuoli
authors.forename: P
authors.initials: P
authors.lastname: Nordera
authors.forename: I
authors.initials: I
authors.lastname: Bernhart
authors.forename: C
authors.initials: C
authors.lastname: Della Volpe
authors.forename: D
authors.initials: D
authors.lastname: Di Giorgio
authors.forename: A
authors.initials: A
authors.lastname: Ballio
authors.forename: G
authors.initials: G
authors.lastname: Menestrina
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D008563:Membrane Lipids / Q000737:chemistry; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry / Q000235:genetics / Q000633:toxicity*; D011549:Pseudomonas / Q000737:chemistry / Q000235:genetics / Q000472:pathogenicity*; D011817:Rabbits; D013261:Sterols / Q000737:chemistry; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D008563:Membrane Lipids; D008567:Membranes, Artificial; D010456:Peptides, Cyclic; D013261:Sterols; C098017:syringopeptin 25-A; C032677:syringomycin; C069036:syringotoxin B