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New technique reveals cross-talk between 2 essential cellular processes
Date:7/19/2012

that the two mechanisms sometimes work in tandem, especially during apoptosis. But those studies had been focused on individual apoptosis-driving enzymes and their biochemical partners, rather than on the "global" apoptosis process within cells.

"In this study, we wanted to develop a global method to let us see all the signs of cross-talk between proteolysis and phosphorylation during apoptosis," said Melissa M. Dix, a research associate in the Cravatt laboratory. Dix was a lead author of the paper, along with then-graduate student Gabriel M. Simon, who is now a postdoctoral researcher at Washington University, St. Louis.

New Insights

Dix and Simon built on an earlier proteolysis-mapping method that they had described in Cell in 2008. Known as PROTOMAP, it can be used to generate a detailed picture of the protein cleavage events in cells during a process of interest. For the new study, the researchers added a technique for detecting phosphorylation events, plus another recently-developed proteomics technology, SILAC, which enables researchers to distinguish, within a given sample, copies of proteins that have come from different cell populations. The researchers then applied the combined techniques to populations of control cells and apoptotic cells, in order to find the proteolysis and phosphorylation events that happened only during apoptosis.

They detected more than 700 apoptosis-specific proteolysis eventsmostly mediated by apoptosis-driving enzymes known as caspasesincluding many that had not been reported before. The new mapping also revealed for the first time an extensive, apoptosis-specific network of phosphorylation events, many of which were clearly connected to proteolysis events. "Just looking at the map of phosphorylation events, we could see that they were unusually common around sites of known caspase cleavage," said Dix.

Previous studies had hinted that the phosphorylation of a protein near one of
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Contact: Mika Ono
mikaono@scripps.edu
Scripps Research Institute
Source:Eurekalert  

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