Yashwanth Ashok

University of Oulu
Supervisor: Veli-Pekka Jaakola
Funding: ISB
Date: 2010-09-10

Biochemical and Biophysical studies on Adenosine A2A receptor and its interacting proteins.

Humans encode four Adenosine receptors (A1, A2A, A2B and A3) all of which belong to the G Protein Coupled Receptor (GPCR) superfamily. Adenosine receptors regulate the physiology of neurotransmitter release, cardiovascular, immune and other major systems in human body. Our study focuses on human Adenosine A2A receptor and its interacting proteins.

The crystal structure of the Adenosine A2A receptor was solved to 2.6 bound to an antagonist. In order to increase the probability of crystallogenesis the flexible third intracellular loop was replaced by T4 lysozyme and carboxyl terminus was also truncated. However the carboxyl terminus interacts with five other interacting proteins that mediate processes such as G protein independent signaling, ER quality control and sustained signaling in plasma membrane. This project aims in solving the structure of the carboxyl terminus of adenosine A2A receptor in complex with interacting proteins. We intend to employ a variety of techniques such as GST pull-down assays, co-immunoprecipitation, size exclusion chromatography, site-directed fluorescence labeling and NMR and X-ray crystallography to analyze interactions between these proteins in addition to other biophysical methods required to understand the energetic basis of interactions.