Supported Solid Lipid Bilayers as a Platform for Single-Molecule Force Measurements
| Autor: | Gero Lutz Hermsdorf, Tobias Jörg Jachowski, Michael Kittelberger, Mohammad Kazem Abdosamadi, Ammara Maqbool, Erik Schäffer, Swathi Sudhakar |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
chemistry.chemical_classification
Chemistry Mechanical Engineering Biomolecule Lipid Bilayers Force spectroscopy Kinesins Bioengineering DNA 02 engineering and technology General Chemistry Microscopy Atomic Force 021001 nanoscience & nanotechnology Condensed Matter Physics Microtubules Mechanobiology Models Chemical Optical tweezers Microscopy Molecular motor Fluorescence microscope Biophysics General Materials Science 0210 nano-technology Lipid bilayer |
| Zdroj: | Nano Letters. 19:8877-8886 |
| ISSN: | 1530-6992 1530-6984 |
| Popis: | Biocompatible surfaces are important for basic and applied research in life science with experiments ranging from the organismal to the single-molecule level. For the latter, examples include the translocation of kinesin motor proteins along microtubule cytoskeletal filaments or the study of DNA-protein interactions. Such experiments often employ single-molecule fluorescence or force microscopy. In particular for force measurements, a key requirement is to prevent nonspecific interactions of biomolecules and force probes with the surface, while providing specific attachments that can sustain loads. Common approaches to reduce nonspecific interactions include supported lipid bilayers or PEGylated surfaces. However, fluid lipid bilayers do not support loads and PEGylation may require harsh chemical surface treatments and have limited reproducibility. Here, we developed and applied a supported solid lipid bilayer (SSLB) as a platform for specific, load bearing attachments with minimal nonspecific interactions. Apart from single-molecule fluorescence measurements, anchoring molecules to lipids in the solid phase enabled us to perform force measurements of molecular motors and overstretch DNA. Furthermore, using a heating laser, we could switch the SSLB to its fluid state allowing for manipulation of anchoring points. The assay had little nonspecific interactions, was robust, reproducible, and time-efficient, and required less hazardous and toxic chemicals for preparation. In the long term, we expect that SSLBs can be widely employed for single-molecule fluorescence microscopy, force spectroscopy, and cellular assays in mechanobiology. |
| Databáze: | OpenAIRE |
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