Study reveals insights into potential target for new medicines

New research efforts have revealed new structural details of a hormone receptor that has puzzled researchers for two decades and prompted them to believe that it could be a target for new medicines related to cardiovascular conditions, neuropathic pain and tissue growth.

The new findings about one of the angiotensin II receptors, called AT2, were made possible by using powerful X-rays from Linac Coherent Light Source (LCLS) at the U.S. Department of Energy's SLAC National Accelerator Laboratory and Argonne National Laboratory's Advanced Photon Source (APS).

AT2's partner, AT1, has been successfully used as a target for high blood pressure medications.

Both AT1 and AT2 are key components of a hormone system that helps regulate blood pressure and sodium levels in the blood.

They are known as "membrane proteins" because they straddle the plasma membranes of cells, where they receive signals from hormones outside the cell and pass them along to soluble partners inside the cell, such as G proteins, setting off a cascade of events that spread the signal cell-wide.

"Unlike its sibling AT1, the AT2 receptor has not been fully understood," said Vadim Cherezov, chemistry professor at the University of Southern California and principal investigator of the study published in Nature.

"Through this study we uncovered many important details about the AT2 receptor structure and how it binds to chemical compounds, information that will stimulate further studies of the receptor's function and could enable an exciting next step in drug discovery."

Many hypertension medications currently on the market target the AT1 receptor because of its well-understood role in blood pressure regulation; they block AT1 to bring blood pressure down.

Cherezov led earlier experiments at LCLS that provided structural details of receptor blockers bound to AT1. The AT2 receptor, on the other hand, is still an elusive drug target despite multiple studies of its function.

Some, but not all, have shown it counteracts the effects of AT1; others point to its potential for tissue protection and regeneration; and still others suggest it could play a role in blocking pain.

"There are no approved drugs yet that act on AT2 receptors, but pharmaceutical companies are actively working on developing compounds that will activate or block these receptors," Cherezov said. "One such compound, called EMA401, is being tested in patients for the treatment of neuropathic pain."

In the latest study, Cherezov's team set out to do two things: find out how AT2 differs from AT1, so they can find ways of selectively activating or blocking it; and better understand why AT2 -- which like AT1 has all the classic features of a G-protein coupled receptor (GPCR) -- fails to activate G-proteins, which spread signals inside cells, or interact with arrestin, which turns those signals off.

In the experiments, the researchers looked at two different kinds of samples, which were formed into crystals for examination with X-rays.

In one, the AT2 receptor was bound to a selective compound, one that binds only to AT2. These crystals were so small that they could only be studied at SLAC's X-ray free-electron laser LCLS, where they were streamed across a beam of ultrafast X-ray pulses.

The results of the experiments were surprising in several ways, according to Cherezov. First, although both compounds were designed to block and deactivate the receptors, they left AT2 in a state that appeared to be active.

But on the inward-facing side of AT2, the site where a G-protein would normally bind and spread the signal was blocked. "This basically explains why these receptors do not activate G-proteins," Cherezov said. "They are activated but self-inhibited."

In addition, although AT1 and AT2 were thought to be very similar, the pockets where the receptors bind to the compounds exhibited marked differences.

"This is something we have never seen with GPCRs," Cherezov was quoted as saying in a news release. "The idea was always that receptors that bind to the same compounds would have very similar pockets, so efforts to develop drugs that act on AT2 started with the same basic structures as drugs that act on AT1. Now we see that we may have to start with entirely different drug-like molecules that are tailored to fit the AT2 receptor, which could set the drug discovery process in a different direction." Endit