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Expression, enzymology and structural determination of the new phosphatase ship2

Objective

The SH2-containig inositol 5-phosphase (SHIP2) is a new PtdInsP3 5-phophatase directly involved in insulin sensitivity . The two main objectives of this project are: a ) obtain recombinant SHIP2 in large amounts to develop a rapid assay of activity b ) determine structural infonnation after cristallography . For the first six months, we would like to produce SHIP2 or the catalytic domain of SHIP2 either in baculo-infected cells or in bacteria. Then, for the next six months, we would like to set up a large scale production of the protein. This recombinant protein will be used on the one hand to develop a rapid activity assay , and on the other hand to permit the cristallisation of SHIP2. So, the second year, we would like to obtain cristals of SHIP2, to do the diffraction of the x- rays. The structural detennination of SHIP2, and more particularly of SHIP2 catalytic site will provide tools for structure-based design of selective inhibitors of SHIP2 phosphatase. And these inhibitors will have potential application for the treatment of type II diabete.

Coordinator

UNIVERSITE LIBRE DE BRUXELLES
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Belgium