title: Interfacial structure of wild-type and mutant forms of puroindoline-b bound to DPPG monolayers.
abstract: The interaction of wild-type puroindoline-b (Pin-b+) and two mutant forms having single residue substitutions (G46S or W44R) with L-alpha-dipalmitoylphosphatidyl-dl-glycerol (DPPG) as a Langmuir monolayer at the air/water interface was investigated by neutron reflectivity (NR) and Brewster angle microscopy (BAM). NR profiles were fitted using a three-layer model to enable differences in penetration of protein between the lipid headgroup and acyl regions to be determined. The data showed similar surface excesses for each of the three proteins at the interface; however, it was revealed that the depth of penetration of protein into the lipid region differed for each protein with Pin-b+ penetrating further into the acyl region of the lipid compared to the mutant forms of the protein that interacted with the headgroup region only. BAM images revealed that the domain structure of the DPPG monolayers was disrupted when Pin-b+ adsorption had reached equilibrium, suggesting protein penetration had led to compression of the lipid region. In contrast, the domain structure was unaffected by the W44R mutant, suggesting no change in compression of the lipid region and hence little or no penetration of protein into the lipid layer.
authors.affiliation: Reading School of Pharmacy and Department of Food Biosciences, University of Reading, P.O. Box 226, Whiteknights, Reading RG6 6AP, United Kingdom.
authors.forename: Luke A
authors.initials: LA
authors.lastname: Clifton
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Green
authors.forename: Arwel V
authors.initials: AV
authors.lastname: Hughes
authors.forename: Richard A
authors.initials: RA
authors.lastname: Frazier
mesh_terms: D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D010715:Phosphatidylglycerols / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011312:Pressure; D011485:Protein Binding; D014908:Triticum / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D010715:Phosphatidylglycerols; D010940:Plant Proteins; C082942:puroindoline protein, Triticum aestivum; C030345:1,2-dipalmitoylphosphatidylglycerol