Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Arman Kulyyassov1, Muhammad Shoaib2, Chloe Robin2, Erlan Ramanculov1 and Vasily Ogryzko2
1National Center for Biotechnology, Kazakhstan 2University of Paris-Sud, France
Posters-Accepted Abstracts: J Proteomics Bioinform
Protein-protein interactions (PPI) play a key role in many processes within the cell and their affinity and specificity are fine-tuned with respect to the functions they perform. Disturbances in the signal transduction pathways associated with PPI lead to the development and progression of cancer. We have used method called the Proximity Utilizing Biotinylation (PUB) based on coexpression within a single cell of the recombinant proteins. The protein of interest fused with biotin ligase BirA and its partner with the biotin acceptor peptide BAP for study of PPI in vivo. The same protein can exist in several complexes in the cell and its properties can vary depending on the proximity partners (PP). One can purify a particular PP-dependent sub-fraction of a BAP-fused protein biochemically and then study its properties, example its post-translational modifications. To test this possibility experimentally, we have analyzed the ubiquitination status of the subset of replication processivity factor PCNA that is located in proximity to the translation polymerase POLH after UV irradiation of cells. UVC irradiation induces ubiquitination of PCNA which then recruits POLH to the sites of DNA damage. The PUB method was also used to study ubiquitin-dependent signaling and regulation. The coexpression of BirA-POLH with the BAP-ubiquitin leads to biotinylation of ubiquitin-modified proteins interacting with the BirAfusion. We showed that the preferential biotinylation of BAP-Ubi-PCNA by the BirA-POLH fusion depends on the integrity of the UBZ and PCNA-binding domains of POLH. Using LC-MS/MS we identified the ubiquitinated proteins interacting with the BirA fusion of interest.
Arman Kulyyassov graduated from the Novosibirsk State University (Russia) in 1992. He has completed his PhD on specialty of bioorganic chemistry. He has published more than 40 papers in international journals related to chemistry of natural compounds. During Post-doctoral studies in the Institut de Cancérologie Gustave Roussy in 2006-2010 (Villejuif, France) he specialized in proteomics, molecular and cell biology. At present he is the leading researcher in the stem cell lab in National Center for biotechnology (Astana, Kazakhstan) and has 5 papers published in peer-reviewed journals on proteomics.
Email: akulyyasov@gmail.com