PT - JOURNAL ARTICLE AU - Yoonji Lee AU - Songmi Kim AU - Sun Choi AU - Changbong Hyeon TI - Ultraslow water-mediated transmembrane interactions regulate the activation of A<sub>2A</sub> adenosine receptor AID - 10.1101/067751 DP - 2016 Jan 01 TA - bioRxiv PG - 067751 4099 - http://biorxiv.org/content/early/2016/08/04/067751.short 4100 - http://biorxiv.org/content/early/2016/08/04/067751.full AB - Water molecules inside G-protein coupled receptor have recently been spotlighted in a series of crystal structures. To decipher the dynamics and functional roles of internal waters in GPCR activity, we studied A2A adenosine receptor using μsec-molecular dynamics simulations. Our study finds that the amount of water flux across the transmembrane (TM) domain varies depending on the receptor state, and that the water molecules of the TM channel in the active state flow three times slower than those in the inactive state. Depending on the location in solvent-protein interface as well as the receptor state, the average residence time of water in each residue varies from psec to nsec. Especially, water molecules, exhibiting ultraslow relaxation ( nsec) in the active state, are found around the microswitch residues that are considered activity hotspots for GPCR function. A continuous allosteric network spanning the TM domain, arising from water-mediated contacts, is unique in the active state, underscoring the importance of slow waters in the GPCR activation.