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Salk scientists crack molecular code regulating neuronal excitability
Date:3/22/2011

citability.

In the new study Slesinger's team confirmed that SNX27 resides in neurons, just below the membrane where active GIRK channels sit. Additional experiments using brain cells manipulated to express abnormally high SNX27 levels showed that cells were less responsive to drugs that activate channels, suggesting that SNX27 waylays membrane-bound GIRKs and blocks their function.

The fact that SNX27 displays a common protein-interaction signature called PDZ domain suggested how SNX27 grabs its partner: GIRKs contain a short, 4-residue sequence that binds to PDZ domains, a recognition motif Slesinger likens to a zip code. But channels similar to GIRKs, called IRKs, displayed an almost identical sequence but were impervious to destruction by SNX27. "We were puzzled by this similarity and swapped the 4-residue code in IRK with the corresponding sequence from GIRK," says Slesinger. Surprisingly, this IRK/GIRK hybrid did not bind SNX27, indicating that the IRK lacked other elements necessary for SNX27 recognition.

To define these new elements, Slesinger consulted with a long-standing collaborator, Senyon Choe, Ph.D., professor in Salk's Structural Biology Laboratory. Choe is an expert on a technique known as X-ray crystallography, used to determine the three-dimensional structure of proteins. The team scrutinized crystallized forms of SNX27 wrapped around the GIRK binding motif to try to visualize where the proteins made contact.

"We observed a binding cleft in the SNX27 PDZ domain and a region that formed another pocket with a lot of positive charges," says Slesinger. "The GIRK fragment lying there had a negative charge upstream of the 4-residue "zip code". That suggested that this second site allowed a previously unknown electrostatic interaction between these two proteins." Therefore, SNX27 may recognize a 6-residue motif, like the "zip plus 4' code.

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Contact: Gina Kirchweger
kirchweger@salk.edu
858-453-4100 x1340
Salk Institute
Source:Eurekalert  

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