Selection and analysis of a DNA aptamer binding α-amanitin from Amanita phalloides

• Authors: Klaudia Muszyńska , Dominika Ostrowska , Filip Bartnicki , Ewa Kowalska , Małgorzata Bodaszewska-Lubaś , Paweł Hermanowicz , Heinz Faulstich , Wojciech Strzałka
• Languages of publication: EN

Abstracts

EN

Mushroom foraging is very popular in some regions of the world. Sometimes toxic and edible mushrooms are mistaken by mushroom collectors, leading to serious human poisoning. The group of mushrooms highly dangerous for human health includes Amanita phalloides. This mushroom produces a toxic octapeptide called α-amanitin which is an inhibitor of nuclear RNA polymerase II. The inhibition of this polymerase results in the abortion of mRNA synthesis. The ingestion of A. phalloides causes liver failure due to the fact that most of the toxin is uptaken by hepatocytes. The hospitalization of poisoned patients involves the removal of the toxin from the digestive tract, its dilution in the circulatory system and the administration of therapeutic adjuvants. Since there is no effective antidote against amanitin poisoning, in this study we developed a DNA aptamer exhibiting specific binding to α-amanitin. This aptamer was selected using the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) method. Next, its ability of toxin removal from aqueous solution was confirmed by pull-down assay. The aptamer region sufficient for α-amanitin binding was determined. Finally, the dissociation constant of the α-amanitin/DNA aptamer complex was calculated.

Keywords

EN

α-amanitin; mushroom poisoning; aptamer; SELEX

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2017
• Volume: 64
• Issue: 3
• Pages: 401-406

Dates

published

2017

received

2017-03-18

revised

2017-05-22

accepted

2017-05-22

(unknown)

2017-08-09

Contributors

author

Klaudia Muszyńska

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Dominika Ostrowska

• Department of Comparative Biochemistry and Bioanalytics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Filip Bartnicki

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Ewa Kowalska

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Małgorzata Bodaszewska-Lubaś

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Paweł Hermanowicz

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

author

Heinz Faulstich

• Bioorganic Research Group, Max Planck Institute for Medical Research, Heidelberg, Germany

author

Wojciech Strzałka

• Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland

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Identifiers

DOI

10.18388/abp.2017_1615