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A.T. Kulyyassov

National Center for Biotechnology under the Science Committee of Ministry of Education and Science of the Republic of Kazakhstan, 13/1 Valikhanov str., Astana, 010000, Kazakhstan 

G.S. Zhubanova

National Center for Biotechnology under the Science Committee of Ministry of Education and Science of the Republic of Kazakhstan, 13/1 Valikhanov str., Astana, 010000, Kazakhstan 

E.M. Ramankulov

National Center for Biotechnology under the Science Committee of Ministry of Education and Science of the Republic of Kazakhstan, 13/1 Valikhanov str., Astana, 010000, Kazakhstan 

V.V. Ogryzko

Institut Gustave Roussy, CNRS UMR8126, 94805, Villejuif, France, 39 Rue Camilles Desmoulin


The development and progression of cancer is accompanied by changes in gene expression which are often can be caused by both genetic and epigenetic alterations and therefore proteins НР1a, b и g are potential oncomarkers.

We have used method, called the Proximity Utilizing Biotinylation (PUB), based on co-expression within a single cell of the recombinant proteins - the protein of interest fused with biotin ligase BirA and his partner with the biotin acceptor peptide BAP, which allows an accurate quantitative assessment of the extent of their interaction in vivo.

The aim of this work is to develop a method for quantifying interactions in vivo of oncomarker proteins HP1a and HP1b.

In experiments on protein expression of BAP and BirA fusions of HP1a, HP1b and Tap54b in HEK293T cells we found elevated levels of biotinylation due to in vivo interaction of homologous proteins BAP-HP1 and BirA-HP1 (BAP-Tap54b and BirA-Tap54b). Signal ratio of heterologous to homologous interaction in all repeated experiments was 0,4 ± 0,14 (for samples containing BAP-HP1a) and 0,32 ± 0,08 (for samples with BAP-HP1b). Qualitative analysis by LC-MS/MS method of the gel fragments corresponding to the immunoblot bands of expressed proteins allowed to identify peptides relevant to BAP, Tap54b,   HP1a and HP1b.


heterochromatin, euchromatin, protein-protein interactions, biotinylation, oncomarkers, biotin ligase, biotin acceptor peptide, plasmids, transient transfection, western blot, mass spectrometry

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