Layer-by-Layer Assembly of Supported Lipid Bilayer Poly‑l‑Lysine Multilayers
Multilayer lipid membranes perform many important functions in biology, such as electrical isolation (myelination of axons), increased surface area for biocatalytic purposes (thylakoid grana and mitochondrial cristae), and sequential processing (golgi cisternae). Here we develop a simple layer-by-la...
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Published in | Biomacromolecules Vol. 17; no. 1; pp. 324 - 335 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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American Chemical Society
11.01.2016
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Abstract | Multilayer lipid membranes perform many important functions in biology, such as electrical isolation (myelination of axons), increased surface area for biocatalytic purposes (thylakoid grana and mitochondrial cristae), and sequential processing (golgi cisternae). Here we develop a simple layer-by-layer methodology to form lipid multilayers via vesicle rupture onto existing supported lipid bilayers (SLBs) using poly l-lysine (PLL) as an electrostatic polymer linker. The assembly process was monitored at the macroscale by quartz crystal microbalance with dissipation (QCM-D) and the nanoscale by atomic force microscopy (AFM) for up to six lipid bilayers. By varying buffer pH and PLL chain length, we show that longer chains (≥300 kDa) at pH 9.0 form thicker polymer supported multilayers, while at low pH and shorter length PLL, we create close packed layers (average lipid bilayers separations of 2.8 and 0.8 nm, respectively). Fluorescence recovery after photobleaching (FRAP) and AFM were used to show that the diffusion of lipid and three different membrane proteins in the multilayered membranes has little dependence on lipid stack number or separation between membranes. These approaches provide a straightforward route to creating the complex membrane structures that are found throughout nature, allowing possible applications in areas such as energy production and biosensing while developing our understanding of the biological processes at play. |
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AbstractList | Multilayer lipid membranes perform many important functions in biology, such as electrical isolation (myelination of axons), increased surface area for biocatalytic purposes (thylakoid grana and mitochondrial cristae), and sequential processing (golgi cisternae). Here we develop a simple layer-by-layer methodology to form lipid multilayers via vesicle rupture onto existing supported lipid bilayers (SLBs) using poly l-lysine (PLL) as an electrostatic polymer linker. The assembly process was monitored at the macroscale by quartz crystal microbalance with dissipation (QCM-D) and the nanoscale by atomic force microscopy (AFM) for up to six lipid bilayers. By varying buffer pH and PLL chain length, we show that longer chains (≥300 kDa) at pH 9.0 form thicker polymer supported multilayers, while at low pH and shorter length PLL, we create close packed layers (average lipid bilayers separations of 2.8 and 0.8 nm, respectively). Fluorescence recovery after photobleaching (FRAP) and AFM were used to show that the diffusion of lipid and three different membrane proteins in the multilayered membranes has little dependence on lipid stack number or separation between membranes. These approaches provide a straightforward route to creating the complex membrane structures that are found throughout nature, allowing possible applications in areas such as energy production and biosensing while developing our understanding of the biological processes at play. |
Author | Richens, Joanna L Catucci, Gianluca Gilardi, Gianfranco Jeuken, Lars J. C Heath, George R O’Shea, Paul Butt, Julea N Sadeghi, Sheila J Li, Mengqiu Polignano, Isabelle L |
AuthorAffiliation | University of Leeds Cell Biophysics Group, Institute of Biophysics, Imaging and Optical Science, School of Life Sciences Centre for Molecular and Structural Biochemistry, School of Biological Sciences, and School of Chemistry University of East Anglia University of Torino School of Biomedical Sciences Life Sciences and Systems Biology University of Nottingham |
AuthorAffiliation_xml | – name: School of Biomedical Sciences – name: Cell Biophysics Group, Institute of Biophysics, Imaging and Optical Science, School of Life Sciences – name: University of Leeds – name: Life Sciences and Systems Biology – name: Centre for Molecular and Structural Biochemistry, School of Biological Sciences, and School of Chemistry – name: University of Torino – name: University of East Anglia – name: University of Nottingham |
Author_xml | – sequence: 1 givenname: George R surname: Heath fullname: Heath, George R – sequence: 2 givenname: Mengqiu surname: Li fullname: Li, Mengqiu – sequence: 3 givenname: Isabelle L surname: Polignano fullname: Polignano, Isabelle L – sequence: 4 givenname: Joanna L surname: Richens fullname: Richens, Joanna L – sequence: 5 givenname: Gianluca surname: Catucci fullname: Catucci, Gianluca – sequence: 6 givenname: Paul surname: O’Shea fullname: O’Shea, Paul – sequence: 7 givenname: Sheila J surname: Sadeghi fullname: Sadeghi, Sheila J – sequence: 8 givenname: Gianfranco surname: Gilardi fullname: Gilardi, Gianfranco – sequence: 9 givenname: Julea N surname: Butt fullname: Butt, Julea N – sequence: 10 givenname: Lars J. C surname: Jeuken fullname: Jeuken, Lars J. C email: L.J.C.Jeuken@leeds.ac.uk |
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SubjectTerms | Lipid Bilayers - chemistry Liposomes - chemical synthesis Membranes - chemistry Microscopy, Atomic Force Microscopy, Fluorescence Polylysine - chemistry Polymers - chemical synthesis Quartz Crystal Microbalance Techniques Static Electricity Surface Properties |
Title | Layer-by-Layer Assembly of Supported Lipid Bilayer Poly‑l‑Lysine Multilayers |
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