"We can make these modifications in just two chemical steps, which is a great advantage," said Alexander Adibekian, a postdoctoral fellow in the Cravatt lab and first author of the new paper. "And despite this technical simplicity, we were able to generate compounds that were extremely potent and selective."
From the 20 compounds the scientists generated this way, they found three with powerful and highly specific inhibitory effects on individual serine hydrolases with many unknown characteristics.
Most of the study's enzyme-inhibition tests were conducted in mouse cell cultures, a more realistic biochemical environment than traditional "test-tube" biochemical preparations; but for one of the group's inhibitor compounds, AA74-1, the scientists extended their inhibition-measurement techniques to animal models, showing that the compound potently blocked the activity of its target serine hydrolase, acyl-peptide hydrolase, or APEH, without significantly affecting other enzymes.
Not much had been known about APEH, but with its inhibitor AA74-1, the team was able to illuminate the enzyme's normal role in the chemical modification of proteins, showing the levels of more than two dozen proteins dropped sharply when APEH was inhibited.
"This was unexpected and unusual," said Adibekian. "But it's what one wants to see with these compoundsstrong enzyme-inhibiting activity in different tissues, at a low dose. And it's the first time this kind of evaluation has been done in both cultured cells and animal tissues."
The Cravatt lab is now using the expanding number of inhibitors that team members have generated so far to study serine hydrolases with pr
|Contact: Mika Ono|
Scripps Research Institute