Proximity-dependent initiation of hybridization chain reaction
| Autor: | Felipe Marques Souza de Oliveira, M Karoliina Hirvonen, Ola Söderberg, Johannes Haybaeck, Masood Kamali-Moghaddam, Linda Arngården, Liza Löf, Gaëlle Cane, Björn Koos, U. Helena Danielson, Manfred Auer, Carl-Magnus Clausson, Vladimir O. Talibov, Nhan T. Pham, Johan Heldin, Axel Klaesson, Karin Grannas |
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| Rok vydání: | 2015 |
| Předmět: |
Medicin och hälsovetenskap
Epidermal Growth Factor/chemistry Oligonucleotides General Physics and Astronomy Sequence (biology) Plasma protein binding Biology 010402 general chemistry Medical and Health Sciences 01 natural sciences Article General Biochemistry Genetics and Molecular Biology Protein–protein interaction Flow cytometry 03 medical and health sciences Nucleic acid thermodynamics medicine Fluorescent Dyes/chemistry Oligonucleotides/chemistry Fluorescent Dyes 030304 developmental biology 0303 health sciences Multidisciplinary Epidermal Growth Factor medicine.diagnostic_test ErbB Receptors/chemistry Oligonucleotide Nucleic Acid Hybridization Kemi General Chemistry Flow Cytometry Molecular biology Fluorescence 0104 chemical sciences 3. Good health ErbB Receptors Chemical Sciences Biophysics Chain reaction Protein Binding |
| Zdroj: | Nature Communications Koos, B, Cane, G, Grannas, K, Löf, L, Arngården, L, Heldin, J, Clausson, C-M, Klaesson, A, Hirvonen, M K, de Oliveira, F M S, Talibov, V O, Pham, N T, Auer, M, Danielson, U H, Haybaeck, J, Kamali-Moghaddam, M & Söderberg, O 2015, ' Proximity-dependent initiation of hybridization chain reaction ', Nature Communications, vol. 6, 7294 . https://doi.org/10.1038/ncomms8294 |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/ncomms8294 |
| Popis: | Sensitive detection of protein interactions and post-translational modifications of native proteins is a challenge for research and diagnostic purposes. A method for this, which could be used in point-of-care devices and high-throughput screening, should be reliable, cost effective and robust. To achieve this, here we design a method (proxHCR) that combines the need for proximal binding with hybridization chain reaction (HCR) for signal amplification. When two oligonucleotide hairpins conjugated to antibodies bind in close proximity, they can be activated to reveal an initiator sequence. This starts a chain reaction of hybridization events between a pair of fluorophore-labelled oligonucleotide hairpins, generating a fluorescent product. In conclusion, we show the applicability of the proxHCR method for the detection of protein interactions and posttranslational modifications in microscopy and flow cytometry. As no enzymes are needed, proxHCR may be an inexpensive and robust alternative to proximity ligation assays. Proximity ligation assays are a sensitive method for detecting protein interactions, but require the addition of enzymes. Here the authors introduce proxHCR, an enzyme-free method of detecting interactions in close proximity by inducing a hybribization chain reaction (HCR) of fluorescently labelled oligonucleotides. |
| Databáze: | OpenAIRE |
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