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Solving a traditional Chinese medicine mystery
Date:3/3/2011

three groups of enzymes that make RNA and found that low doses of triptolide blocked only one, RNAPII.

But the RNAPII enzyme complex actually requires the assistance of several smaller clusters of proteins, according to Liu, which required more investigative narrowing down. Using a small gene fragment in a test tube, the researchers mixed in RNAPII components and in some tubes included triptolide and some not to see which combinations resulted in manufacture of new RNA. Every combination of proteins that included a cluster called TFIIH stopped working in the presence of triptolide.

But again, TFIIH is made of 10 individual proteins, many of which, according to Liu, have distinct and testable activities. Using information already known about these proteins and testing the rest to see if triptolide would alter their behaviors, the research team finally found that triptolide directly binds to and blocks the enzymatic activity of one of the 10, the XPB protein.

"We were fairly certain it was XPB because other researchers had found triptolide to bind to an unknown protein of the same size, but they weren't able to identify it," says Liu. "

To convince themselves that the interaction between triplotide and XPB is what stops cells from growing, the researchers made 12 chemicals related to triplotide with a wide range of activity and treated HeLa cells with each of the 12 chemicals at several different doses. By both counting cells and testing XPB activity levels, the team found that the two correlate; chemicals that were better at decreasing XPB activity were also better at stopping cell growth and vice versa.

"Triptolide's general ability to stop RNAPII activity explains its anti-inflammatory and anticancer effects," says Liu. "And its behavior has important additional implications for circumventing the resistance that some cancer cells develop to certain anticancer drugs. We're eager to study it further to see what it can
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Contact: Audrey Huang
audrey@jhmi.edu
410-614-5105
Johns Hopkins Medical Institutions
Source:Eurekalert

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