Protein complementation approach for quantitative and qualitative analysis of hTERP in cell

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Abstract

The study of protein functioning in a cell cannot be performed by analyzing their content in the cell. The most commonly used method of analysis, western blotting, cannot always be used due to the impossibility of obtaining antibodies that specifically recognize the protein of interest. At the same time, western blotting is a semi-quantitative method of analysis and does not allow determining the exact content of protein in the cell. In this work, we used the protein complementation method for the qualitative and quantitative assessment of the hTERP protein content in the cell. Using genome editing, a HiBiT appendage was introduced to the C-terminus of the hTERP protein, the complementation of which with LgBiT restores active luciferase. As a result, we determined the hTERP protein content in the HEK293T cell line.

About the authors

M. A. Shamonova

Lomonosov Moscow State University, Department of Chemistry

Email: mprubtsova@gmail.com
Moscow, Russian Federation

M. S. Koriagina

Lomonosov Moscow State University, Department of Chemistry; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: mprubtsova@gmail.com
Moscow, Russian Federation; Moscow, Russian Federation

V. L. Shliapina

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: mprubtsova@gmail.com
Moscow, Russian Federation

O. A. Dontsova

Lomonosov Moscow State University, Department of Chemistry; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences; A. N. Belozersky Institute of Physico-Chemical Biology; Skolkovo Institute of Science and Technology, Center for Molecular and Cellular Biology

Author for correspondence.
Email: mprubtsova@gmail.com
Moscow, Russian Federation; Moscow, Russian Federation; Moscow, Russian Federation; Moscow, Russian Federation

M. P. Rubtsova

Lomonosov Moscow State University, Department of Chemistry; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Email: mprubtsova@gmail.com
Moscow, Russian Federation; Moscow, Russian Federation

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