Protein analysis by time-resolved measurements with an electro-switchable DNA chip
| Autor: | Ulrich Rant, Thomas Welte, Paul A. Hampel, Andreas Langer, Ralf Strasser, Jelena Knezevic, Simone Jähner, Wolfgang Kaiser, Makiko Maruyama, Valentina Villa, Frank Fischer, Matej Svejda |
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| Rok vydání: | 2013 |
| Předmět: |
Protein Folding
Time Factors Materials science General Physics and Astronomy Nanotechnology Molecular Dynamics Simulation Chorionic Gonadotropin Article General Biochemistry Genetics and Molecular Biology Fungal Proteins Molecular dynamics Bacterial Proteins Electricity Humans Molecule Oligonucleotide Array Sequence Analysis Fungal protein Multidisciplinary Tethering DNA General Chemistry Protein tertiary structure Protein Structure Tertiary ddc Characterization (materials science) Folding (chemistry) Kinetics Chemical physics Immunoglobulin G Protein folding Protein Processing Post-Translational |
| Zdroj: | Nature Communications |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/ncomms3099 |
| Popis: | Measurements in stationary or mobile phases are fundamental principles in protein analysis. Although the immobilization of molecules on solid supports allows for the parallel analysis of interactions, properties like size or shape are usually inferred from the molecular mobility under the influence of external forces. However, as these principles are mutually exclusive, a comprehensive characterization of proteins usually involves a multi-step workflow. Here we show how these measurement modalities can be reconciled by tethering proteins to a surface via dynamically actuated nanolevers. Short DNA strands, which are switched by alternating electric fields, are employed as capture probes to bind target proteins. By swaying the proteins over nanometre amplitudes and comparing their motional dynamics to a theoretical model, the protein diameter can be quantified with Angström accuracy. Alterations in the tertiary protein structure (folding) and conformational changes are readily detected, and even post-translational modifications are revealed by time-resolved molecular dynamics measurements. The comprehensive bioanalysis of proteins usually requires multi-step surface and mobile phase measurements. Here, the authors use chips functionalized with dynamically actuated nanolevers—DNA strands that can be switched in an electric field—to obtain motional dynamic measurements of proteins on a chip. |
| Databáze: | OpenAIRE |
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