Structure/Function of Protein Tyrosine Phosphatases

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DESCRIPTION (provided by applicant): The long-term objectives of this research program are to characterize the structure and function of protein tyrosine phosphatases (PTPs). The PTPs constitute a large family of signaling enzymes that together with protein tyrosine kinases (PTKs) modulate the cellular level of tyrosine phosphorylation. Disturbance of the normal balance between PTK and PTP activity results in aberrant tyrosine phosphorylation, which has been linked to the etiology of several human diseases, including cancer. Thus, a complete understanding of the physiological roles of protein tyrosine phosphorylation and how this process is deregulated in human diseases must necessarily encompass the characterization of PTPs. Such understanding may lead to the development of novel therapeutics that selectively target elements of signaling pathways for the treatment of human diseases. This competitive renewal focuses on SHP2 (Src homology 2 (SH2)-domain containing protein tyrosine phosphatase-2), which is the first bona fide oncoprotein identified in the PTP superfamily. SHP2 is ubiquitously expressed and positively regulates signaling from receptor tyrosine kinases through the activation of the Ras/ERK1/2 cascade. Consistent with its oncogenic role, germline autosomal dominant SHP2 mutations cause clinically similar LEOPARD syndrome (LS) and Noonan syndrome (NS), both of which are associated with increased risk of malignancy. In addition, somatic SHP2 mutations contribute to many forms of leukemia and solid tumors. However, although SHP2 mutations are associated with a number of developmental and neoplastic disorders, it remains unclear how SHP2 mutations alter cellular signaling to produce disease phenotypes. For example, NS or neoplasia-associated SHP2 mutants are constitutively active, resulting in gain-of-function effects. In contrast, mutations associated with LS reduce SHP2 phosphatase activity. These findings generated an enigma: how do SHP2 mutations with opposite effects elicit overlapping phenotypes? We hypothesize that pathogenic SHP2 mutations alter not only SHP2 phosphatase activity but also its molecular switching mechanism to drive disease outcomes and thus detailed understanding of the structure and function of SHP2 will reveal critical signaling events that underlie the diseases. The goals of this project ar to understand the molecular basis of disease-associated SHP2 mutations and to define the chain of molecular events coupling SHP2 dysfunction to the various LS abnormalities. A multidisciplinary approach, involving innovative combinations of X-ray crystallography, mass spectrometry, combinatorial chemistry, site-directed mutagenesis, enzyme kinetics, and cell biology will be employed to: 1) characterize the structural and biochemical properties of the LS mutants, and 2) define the signaling mechanisms mediated by the LS mutants. Successful completion of this project will create a solid framework for understanding how individual SHP2 mutations cause diseases and provide insight into novel points of therapeutic intervention for these diseases.