A significative part of world population suffers whit thyroid related problems. Thyroid hormone regulates a lot of functions from endocrine system by its interaction with thyroid hormone receptor (TR). Thyroid hormone presence induces a conformational exchange in its receptor, TR, which leads to the interaction with a coactivator protein like GRIP, for example. TR (association) with coactivator helps in chromatin opening through histone acetylases leading to transcription activation of several genes, mainly of genes related to proteins wich regulate basal metabolism.This project aims to study nuclear receptor-coactivators complex formation and characterization to map its interfaces of interaction. To reach this objective it will be executed assays to evaluate complex stoichiometry, binding affinity among these proteins and also the ligand role in these receptors.Proteins will be purified and complexes will be formed by pull down like assays or coexpression. Complex will be analysed by mass spechometry using cross-linking reaction to map the interaction among NR-CoA, looking for to identify new interactions. Complex stoichiometry will be investigated through analytical ultracentrifugation; the binding affinity among the proteins will be executed applying fluorescence anisotropy. Finally, it will be executed crystallization assays of complex of complex which will bring a better understanding about their structures. It's important to mention that knowing the interactions among these proteins will give subsidy for better compreention about TR action mechanism. This is direct related to drug design once the surface of interaction between two proteins could be considered as a new target for ligands activation.
News published in Agência FAPESP Newsletter about the scholarship: