Isolation of High-Molecular-Weight Activation Complexes of Initiator Caspases in DNA Damage
- Autores: Zamaraev A.1, Egorshina A.1, Lavrik I.1,2, Zhivotovsky B.1,3, Kopeina G.1
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Afiliações:
- Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University
- Otto von Guericke University
- Karolinska Institutet
- Edição: Volume 168, Nº 1 (2019)
- Páginas: 132-140
- Seção: Translated from Kletochnye Tekhnologii v Biologii i Meditsine (Cell Technologies in Biology and Medicine)
- URL: https://journals.rcsi.science/0007-4888/article/view/242283
- DOI: https://doi.org/10.1007/s10517-019-04663-1
- ID: 242283
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Resumo
Initiation of apoptosis by chemotherapeutic drugs is one of the most effective approaches to the treatment of cancers. Caspases, the main enzymes of apoptosis, undergo activation to initiate cell death. Activation of initiator caspases requires their binding to special protein complexes. For elucidation of the mechanisms of apoptosis, these complexes should be isolated. However, their purification is challenging because they are formed in the cell in negligible amounts and rapidly degrade. We have developed an effective way to isolate caspase activation complexes formed in tumor cells in response to DNA damage. The method is based on combination of gel filtration with immunoprecipitation. The first stage is aimed at the separation of the high-molecular-weight caspase activation complexes and their monomeric forms, which allows increasing the efficiency of isolation of complexes at the second stage.
Sobre autores
A. Zamaraev
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University
Email: lirroster@gmail.com
Rússia, Moscow
A. Egorshina
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University
Email: lirroster@gmail.com
Rússia, Moscow
I. Lavrik
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University; Otto von Guericke University
Email: lirroster@gmail.com
Rússia, Moscow; Magdeburg
B. Zhivotovsky
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University; Karolinska Institutet
Autor responsável pela correspondência
Email: boris.zhivotovsky@ki.se
Rússia, Moscow; Stockholm
G. Kopeina
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University
Autor responsável pela correspondência
Email: lirroster@gmail.com
Rússia, Moscow