Scientists have examined deep into the heart of a key protein used in drug design and discovered dynamic structural features that may lead to new ways to target diseases. A protein called A2A adenosine receptor (A2aAR), is a member of the G-protein-coupled receptor (GPCR) family, which are the targets of roughly 40 percent of all approved pharmaceuticals.
The new, more detailed image of A2aAR's signaling mechanism reveals key parts of its inner workings, including an amino acid that acts like a "toggle switch" to control signaling across the cell membrane. This study was published in the journal Cell.
A new study, the researchers aimed to better understand the relationship between A2aAR function and dynamic changes in its structure to help inform drug design. All human cells contain A2aAR and other GPCRs embedded in their plasma membrane. More than 800 GPCRs have been discovered in the human body, and each has a role in regulating a bodily function.
The research built on previous studies where scientists used an imaging technique called x-ray crystallography to determine A2aAR's three-dimensional structure. The images showed that A2aAR looks like a chain that crisscrosses the cell membrane and has an opening on the side facing out of the cell. The region of the GPCR structure that sticks out of the membrane interacts with drugs and other molecules to signal to partner proteins inside the cell.
Although crystal structures provided a key outline of the receptor's shape in inactive and active-like states, they could not show motion and changes in structure when A2aAR meets new binding partners, such as pharmaceutical candidates. In short, the researchers in the new study needed to investigate why A2aAR works the way it does. To solve this problem, the researchers used a technique called nuclear magnetic resonance (NMR) spectroscopy, which creates strong magnetic fields to locate the positions of probes in a sample.
With NMR, scientists can determine the structures of proteins and study their dynamic properties in solution at physiological temperatures — the way they exist in the human body. Researchers follow the effects of drug binding at the extracellular surface on changes in protein structure and dynamics at the intracellular surface, the structural basis of signal transfer across the heart of the GPCR. Two details in A2aAR's structure gave researchers insight into how future drugs could manipulate the receptor. One key finding was that replacing one amino acid in the receptor's center destroyed the receptor's ability to send signals into the cell.
The researchers also revealed the activity of a "toggle switch" in A2aAR. Previous studies suggested that one of the tryptophan amino acids in A2aAR flips up and down in concert with A2aAR's activity. With NMR, the scientists directly observed this unique tryptophan as it changed orientations in response to different drugs. Chemists could potentially modify drugs to manipulate this switch and control A2aAR signaling.