Ariad Pharmaceuticals, Inc.

Mark J Zoller

Dennis Alan Holt

Anthony J Pawson

Ian MacNeil

Berkley A Lynch

Jay Morganstern

Jennifer L Karas

Marcos H Hatada

Joan Brugge

Xiaode Lu

Lily Shiue

Ellen R Laird

Lynne Zydowsky

The invention relates to human ZAP-70, and in particular, to the region of ZAP-70 containing the tandem Src homology-2 (&ldquo;SH2&rdquo;) domains, to crystalline forms thereof, liganded or unliganded, which are particularly useful for the determination of the three-dimensional structure of the protein. The three dimensional structure of the tandem SH2 region of ZAP provides information useful for the design of pharmaceutical compositions which inhibit the biological function of ZAP and other members of the ZAP family of SH2 domain-containing proteins, particularly those biological functions mediated by molecular interactions involving one or both SH2 domains.

Three dimensional structure of a ZAP tyrosine protein kinase fragment and modeling methods

An in vitro assay method permits the identification of a test substance which inhibits the mutual association of two molecules. The method includes the steps of providing two components capable of mutual association, one of said components bearing a covalently linked fluorophore; preparing a mixture containing the two components and at least one test substance; irradiating the mixture with polarized light of a suitable wavelength permitting excitation of the fluorophore as indicated by emission of polarized light; measuring the degree of polarization of the emission, and determining the effect of the presence or concentration of the test substance in decreasing the observed emission polarization of a mixture of the two components alone. Inhibitory activity of the test substance correlates with decreased depolarization values.

In vitro fluorescence polarization assay

Human syk

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Weston, MA, United States of America

Mark J Zoller