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Toxic compound opens potentially important cell gates

Andrew Scharenberg's lab at Harvard Medical School, which specializes in calcium channels' roles in the immune system, identified two genes that they thought were linked to calcium channels.

Scharenberg noted that both genes contained a section of DNA similar to a DNA segment that is the defining characteristic of a family of genes Bessman's group discovered and calls "Nudix" hydrolases. The characteristic section of these genes is known as a "Nudix box."

"We think this family of genes has been around for a long time," says Christopher Dunn, a research technician at Hopkins and an author on the "Nature" paper. "Although the proteins we worked with in this particular project are from humans, the Nudix hydrolases are found in the most primitive to the highest organisms, with nearly 600 members in 200 species identified so far."

Cells most commonly use Nudix proteins as "housecleaners"that break apart potentially harmful derivatives of nucleoside diphosphates, but Bessman's group has also shown Nudix proteins have been adapted to a variety of other functions.

Scharenberg sent clones of the Nudix genes he'd found to Bessman, and Dunn transferred the shorter gene into E. coli bacteria. When the bacteria used the gene to make proteins, Dunn tested them against a series of compounds that bind to Nudix boxes. The protein would only bind to ADP-ribose, a toxic biochemical that is used by some organisms including diptheria to disable cells.

"ADP-ribose is also involved in apoptosis, a self-destruct mechanism used by sick or genetically damaged cells," says Dunn. "Free ADP-ribose is released all the time in healthy cells as a byproduct of other reactions, but it's always cleaned up."

The second, larger gene identified by Scharenberg had a Nudix box at one end. When Bessman and Dunn isolated that segment of the larger protein and repeated the experiment they'd performed on the shorter gene, they found that it also would only bind to ADP-ribose.

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Contact: Michael Purdy
mcp@jhu.edu
410-516-7160
Johns Hopkins University
5-Jun-2001


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