2016
DOI: 10.3791/53373
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Membrane Transport Processes Analyzed by a Highly Parallel Nanopore Chip System at Single Protein Resolution

Abstract: Membrane protein transport on the single protein level still evades detailed analysis, if the substrate translocated is non-electrogenic. Considerable efforts have been made in this field, but techniques enabling automated high-throughput transport analysis in combination with solvent-free lipid bilayer techniques required for the analysis of membrane transporters are rare. This class of transporters however is crucial in cell homeostasis and therefore a key target in drug development and methodologies to gain… Show more

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Cited by 2 publications
(2 citation statements)
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“…LUVs composed of 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphoethanolamine (POPE), and 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphoglycerol (POPG) were purchased from Avanti Polar Lipids Inc. (Hamburg, Germany) and mixed in molar ratios of 4:3:3 to a concentration of 5 mg/mL and subsequently resolved in HEPES buffer (20 mM HEPES/NaOH, 150 mM NaCl, pH 7.4). LUV formation was conducted as described. , Liposomes were extruded 21 times through 100 nm polycarbonate membranes at the LiposoFast-Basic extruder (AVESTIN, Mannheim, Germany). Vesicle sizes were analyzed using the nanoparticle tracking system NanoSight LM10 (Malvern, Herrenberg, Germany).…”
Section: Methodsmentioning
confidence: 99%
“…LUVs composed of 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphoethanolamine (POPE), and 1-palmitoyl-2-oleoyl- sn -glycero-3-phosphoglycerol (POPG) were purchased from Avanti Polar Lipids Inc. (Hamburg, Germany) and mixed in molar ratios of 4:3:3 to a concentration of 5 mg/mL and subsequently resolved in HEPES buffer (20 mM HEPES/NaOH, 150 mM NaCl, pH 7.4). LUV formation was conducted as described. , Liposomes were extruded 21 times through 100 nm polycarbonate membranes at the LiposoFast-Basic extruder (AVESTIN, Mannheim, Germany). Vesicle sizes were analyzed using the nanoparticle tracking system NanoSight LM10 (Malvern, Herrenberg, Germany).…”
Section: Methodsmentioning
confidence: 99%
“…Nanospot GmbH′ developed an array of nanopores utilizing reactive ion etching techniques to investigate fusion process and membrane transport . In another approach, an array of nanopores was fabricated in prepattern arranged silicon cavities. The hybrid biological solid-state nanopore has tremendous applications to study sequencing of DNA, and bacteriophage phi29 connector channel fused in bilayer has revealed the dynamics of ds DNA translocation. , …”
Section: Introductionmentioning
confidence: 99%