title: Time influence on the interaction between Cyt2Aa2 and lipid/cholesterol bilayers.
abstract: Protein-membrane interactions are still an important topic of investigation. One of the suitable experimental techniques used by the scientific community to address such question is atomic force microscopy. In a previous work, we have reported that the binding mechanism between the cytolytic and antimicrobial protein (Cyt2Aa2) and lipid/cholesterol bilayers was concentration-dependent, leading to either the formation of holes in the bilayer or aggregates. Here we study such binding mechanism as a function of time at low protein concentrations (10 µg/mL). We demonstrate that although holes are formed during the first stages of the protein-lipid interaction, a reparation process due to molecular mobility in the bilayer leads to a homogenous and isotropic protein-lipid/cholesterol layer within 3 hr. The combination of imaging, force spectroscopy, and phase contrast delivered information about topography dynamics (molecular mobility), layer thickness, and mechanical properties of the protein-lipid/cholesterol system. These results highlight the importance of the observation time in (such type of) protein-lipid interactions (at low protein concentrations).
authors.affiliation: Institute for Biophysics, Department of Nanobiotechnology, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria.
authors.forename: Alberto
authors.initials: A
authors.lastname: Moreno-Cencerrado
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya Campus, Nakhon Pathom, 73170, Thailand.
authors.forename: Sudarat
authors.initials: S
authors.lastname: Tharad
authors.affiliation: Institute for Biophysics, Department of Nanobiotechnology, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria.
authors.forename: Jagoba
authors.initials: J
authors.lastname: Iturri
authors.affiliation: National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand.
authors.forename: Boonhiang
authors.initials: B
authors.lastname: Promdonkoy
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya Campus, Nakhon Pathom, 73170, Thailand.
authors.forename: Chartchai
authors.initials: C
authors.lastname: Krittanai
authors.affiliation: Institute for Biophysics, Department of Nanobiotechnology, University of Natural Resources and Life Sciences Vienna (BOKU), Muthgasse 11, Vienna, 1190, Austria. jose.toca-herrera@boku.ac.at.
authors.forename: José L
authors.initials: JL
authors.lastname: Toca-Herrera
mesh_terms: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism*; D004731:Endotoxins / Q000737:chemistry* / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D002784:Cholesterol
keywords: atomic force microscopy; binding mechanism; cholesterol-lipid bilayer; cytolytic protein