Use of proximity utilizing biotinylation (PUB) to study protein-protein interactions in vivo
Journal of Proteomics & Bioinformatics

Journal of Proteomics & Bioinformatics
Open Access

ISSN: 0974-276X

Use of proximity utilizing biotinylation (PUB) to study protein-protein interactions in vivo

6th International Conference & Expo on Proteomics

March 29-31, 2016 Atlanta, USA

Arman Kulyyassov and Vasily Ogryzko

National Center for Biotechnology, Kazakhstan
Institut de Canc√?¬?√?¬©rologie Gustave Roussy, France

Posters & Accepted Abstracts: J Proteomics Bioinform

Abstract :

Many cellular processes are carried out by physically interacting proteins and about 80% of them form multi-protein complexes. Identification of protein-protein interactions (PPI) is therefore a critical step in understanding of cells function. We have developed a systematic proteomics-based approach to study PPI which uses Proximity Utilizing Biotinylation (PUB). The coexpression of a protein of interest A, fused to BirA ligase with the fusion of a protein B with BAP (Biotin Acceptor Peptide, specifically biotinylated by BirA) leads to biotinylation of proteins interacting with (or else in proximity to) the BirA-fusion in vivo. Using this method based on enzyme/substrate pair labeling (BirA-A+BAP-B) we demonstrated on several experimental models that the biotinylation is interaction/proximity dependent. Here A and B are next nuclear proteins used in experiments such as paralogues of heterochromatin protein HP1, HP1, HP1, wild type and mutants of transcription factor Kap1, translesion DNA polymerase PolH and E3 ubiquitin ligase RAD18, Proliferative Cell Nuclear Antigen (PCNA), ubiquitin Ub, SUMO-2/3, different types and isoforms of histones H2A, H2Az, H3.1, H3.3, CenpA, H2A.BBD and macroH2A.

Biography :

Email: [email protected]