title: Selected wheat seed defense proteins exhibit competitive binding to model microbial lipid interfaces.
abstract: Puroindolines (Pins) and purothionins (Pths) are basic, amphiphilic, cysteine-rich wheat proteins that play a role in plant defense against microbial pathogens. This study examined the co-adsorption and sequential addition of Pins (Pin-a, Pin-b, and a mutant form of Pin-b with Trp-44 to Arg-44 substitution) and β-purothionin (β-Pth) model anionic lipid layers using a combination of surface pressure measurements, external reflection FTIR spectroscopy, and neutron reflectometry. Results highlighted differences in the protein binding mechanisms and in the competitive binding and penetration of lipid layers between respective Pins and β-Pth. Pin-a formed a blanket-like layer of protein below the lipid surface that resulted in the reduction or inhibition of β-Pth penetration of the lipid layer. Wild-type Pin-b participated in co-operative binding with β-Pth, whereas the mutant Pin-b did not bind to the lipid layer in the presence of β-Pth. The results provide further insight into the role of hydrophobic and cationic amino acid residues in antimicrobial activity.
authors.affiliation: Reading School of Pharmacy, University of Reading, Whiteknights, Reading, United Kingdom.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Sanders
authors.forename: Luke A
authors.initials: LA
authors.lastname: Clifton
authors.forename: Cameron
authors.initials: C
authors.lastname: Neylon
authors.forename: Richard A
authors.initials: RA
authors.lastname: Frazier
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Green
mesh_terms: D000327:Adsorption; D000890:Anti-Infective Agents / Q000378:metabolism; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D001419:Bacteria / Q000737:chemistry*; D001667:Binding, Competitive; D008563:Membrane Lipids / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D014908:Triticum / Q000737:chemistry*
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D008563:Membrane Lipids; D010940:Plant Proteins; C082942:puroindoline protein, Triticum aestivum; C016085:purothionin