Purification and concentration of RNA polymerase on Ni-lipid monolayers
- Authors: Karlova M.G.1, Volokh O.I.1, Chertkov O.V.1, Kirpichnikov M.P.1,2, Studitsky V.M.1,3, Sokolova O.S.1
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Affiliations:
- Department of Bioengineering, School of Biology
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Fox Chase Cancer Center
- Issue: Vol 43, No 6 (2017)
- Pages: 658-663
- Section: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228728
- DOI: https://doi.org/10.1134/S1068162017060048
- ID: 228728
Cite item
Abstract
Knowledge of the chromatin transcription intermediate structures can shed light on the functioning of RNA polymerase in the cell, which is important both for basic science and for the development of therapeutic approaches for the treatment of diseases associated with transcription disorders. In this study, we used affinity monolayers formed by lipids bound to Ni to concentrate RNA polymerase from a dilute solution containing glycerol. It has been shown that RNA polymerase can be isolated and concentrated on a Ni-lipid monolayer from solutions containing cryoprotectants, which usually make it difficult to visualize samples by electron microscopy. This method of isolation allowed us to obtain more diverse spatial orientations of molecules on carbon film, which facilitates three-dimensional reconstruction from TEM data. The results obtained can be used in further structural studies of the transcription processes to identify interactions in transcriptional complexes containing RNA polymerase.
About the authors
M. G. Karlova
Department of Bioengineering, School of Biology
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234
O. I. Volokh
Department of Bioengineering, School of Biology
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234
O. V. Chertkov
Department of Bioengineering, School of Biology
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234
M. P. Kirpichnikov
Department of Bioengineering, School of Biology; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234; Moscow, 117437
V. M. Studitsky
Department of Bioengineering, School of Biology; Fox Chase Cancer Center
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234; Philadelphia, PA, 19111
O. S. Sokolova
Department of Bioengineering, School of Biology
Author for correspondence.
Email: sokolova@mail.bio.msu.ru
Russian Federation, Moscow, 119234