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Cancer researchers find key protein link
Date:3/27/2014

1 groove formed between the beta cap and iron-sulfur cluster binding domains; the strongest coupling is at the cluster binding domain and some contacts of interest are at the top of the beta-cap domain. Since the iron-sulfur cluster is the functional entity involved in NAF-1 activity, these findings clearly indicate that Bcl-2 interaction with NAF-1 affects its activity, Onuchic said.

The research team used a unique combination of experimental and theoretical methods, including peptide array binding studies with fragments of Bcl-2 to NAF-1; the researchers performed functional studies of cluster transfer and other full-length protein interactions with a spectrometer sensitive to hydrogen/deuterium exchange. They combined their results with a computer-based process created at CTBP called direct coupling analysis (DCA), through which interactions between proteins can be predicted by their genomic roots.

"Each of the three techniques not only confirmed the results of the other methods but also provided unique insights in their own right," said Patricia Jennings, a lead author and CTBP affiliate based at the University of California, San Diego (UCSD), where she is a professor of chemistry and biochemistry.

Jennings said the combined techniques are applicable to biomolecular interactions in general. "DCA helps us efficiently filter through massive amounts of data and does not require high-resolution structural studies, although those are desirable," she said. "Peptide array is powerful for localizing fragments that bind with high affinity, and hydrogen/deuterium exchange studies allow us to monitor parts of the intact protein that are not seen in structural studies and are not amenable to DCA analysis.

"Together, the techniques provide an exquisite synergy," she said.

Jennings is a co-lead author of the study with Onuchic; Rachel Nechushtai, a professor at the Alexander Siberman Life Science Institute of the Hebrew University
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Contact: David Ruth
david@rice.edu
713-348-6327
Rice University
Source:Eurekalert  

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