Production of unstable proteins through the formation of stable core complexes
To assemble stable functional complexes, various partners (multi-domains proteins (X-Y-Z) connected by unstructured linkers (blue lines) allowing high inter domain flexibility (double arrow), single domain proteins (A, B), nucleic acids (DNA or RNA), small molecules (yellow disks)) are necessary.
Nat Commun March 17, 2016
March 29, 2016
The team of Marc Ruff works on chromatin stability and DNA mobility. Scientists have recently developed a methodology that enables the production of stable complexes for their structural and functional studies. These results were published in the journal Nature Communications on March 17, 2016.
The main focus of the team of Marc Ruff is to understand the molecular processes performed by nucleoprotein complexes involving the human immunodeficiency virus (HIV) integrase. The purpose is to provide four dimensional views of these complexes during the replication cycle of HIV.
Towards the establishment of a methodology for stable complex production
Recent advances in biology have shown that most cellular functions are performed by transient multi-protein complexes. Protein purification is a series of processes intended to isolate one or a few proteins from a complex mixture, usually cells or tissues. Protein purification is vital in basic research for the characterization of the function, structure and interactions of the relevant protein. It is also essential in applied research and industry to prepare products and reagents.
The purification of proteins that participate in large transient complexes is impeded by low amounts, heterogeneity, instability and poor solubility. To circumvent these difficulties researchers have set up a methodology that enables the production of stable complexes for structural and functional studies. This procedure is benchmarked and applied to two challenging protein families: the human steroid nuclear receptors and the HIV-1 pre-integration complex.
Therapeutic application of this technology is also considered by the research team. Indeed, the multi-protein complexes obtained are used for the discovery of a new family of medicine: conformational inhibitors. These molecules act by blocking the protein in a given form and will allow the inhibition of the viral infection in the case of HIV.