Abstract
According to the World Health Organization Report, depressive disorders affect about 10% of the population. The molecular mechanism of the pathogenesis of depression is still not well understood. The new findings point to phosphatases as potential targets for effective depression therapy. The aim of the project is the development of method that would enable the identification of Mitogen-Activated Protein Kinase Phosphatase-1 (MKP-1) protein partners using proteomic techniques. The research was carried out using the PC12 cell line, often used as a model for neurobiological research. The use of the procedure for efficient purification of protein complexes – Tandem Affinity Purification (TAP) will facilitate the identification of proteins interacting with MKP-1, a potential goal of effective antidepressant therapy. The presented protocol for purification of protein complexes is universal and can be successfully used in different mammalian cell lines. Identified proteins belong to various groups: cytoskeletal, ribosomal, nucleic acid binding, chaperones, enzymes and may potentially be involved in the molecular mechanism of depression.
Sequential Peptide Affinity Purification System for the Systematic Isolation and Identification of Protein Complexes from Escherichia coli
