Final published version
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Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Layer-by-layer assembly of supported lipid bilayer poly-l-lysine multilayers
AU - Heath, George R.
AU - Li, Mengqiu
AU - Polignano, Isabelle L.
AU - Richens, Joanna L.
AU - Catucci, Gianluca
AU - O'Shea, Paul
AU - Sadeghi, Sheila J.
AU - Gilardi, Gianfranco
AU - Butt, Julea N.
AU - Jeuken, Lars J.C.
PY - 2016/1/11
Y1 - 2016/1/11
N2 - Multilayer lipid membranes form 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 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, whilst at low pH and shorter length PLL, we create close packed layers (average lipid bilayers separations of 2.8 nm 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 whilst developing our understanding of the biological processes at play.
AB - Multilayer lipid membranes form 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 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, whilst at low pH and shorter length PLL, we create close packed layers (average lipid bilayers separations of 2.8 nm 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 whilst developing our understanding of the biological processes at play.
U2 - 10.1021/acs.biomac.5b01434
DO - 10.1021/acs.biomac.5b01434
M3 - Journal article
C2 - 26642374
VL - 17
SP - 324
EP - 335
JO - Biomacromolecules
JF - Biomacromolecules
SN - 1525-7797
IS - 1
ER -