MARS · Mechanism of allosteric regulation of SHP2 phosphatase and its role in cancer and genetic diseases: a multidisciplinary computational, structural and biological approach
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2017-04-01 → 2019-03-31
- EU contribution
- €159,461
- Participants
- 1
- Scheme
- MSCA-IF
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Results in brief
Mechanism of allosteric regulation of SHP2 phosphatase and its role in cancer and geneticdiseases: a multidisciplinary computational, structural and biological approach
Protein kinases and protein phosphatases are involved in the regulation of any kind of cellular process, including protein synthesis, signal transduction, cell division, cellular growth, development and aging. Notwithstanding phosphatases are generally considered as enzymes that downregulate cellular processes, the SH2 domain-containing phosphatase SHP2 is an exception, as it plays a relevant role in the upregulating the signaling cascade in RAS/MAPK pathway. Such a pathway is essential for the cellular development as it controls the cellular growth, homeostasis, motility and apoptosis. Pathogenetic mutations of SHP2 cause severe developmental disorders (i.e., Noonan syndrome and LEOPARD syndrome), and childhood malignancies. The structure of SHP2 includes two Src homology 2 domains, called N-SH2 and C-SH2, followed by the catalytic PTP domain, and a C-terminal tail with a still uncharacterized function. SH2 domains are recognition elements that bind protein sequences or peptides containing a phosphorylated tyrosine (pY); in SHP2, they mediate association to receptor tyrosine kinases, cytokine receptors and scaffolding adaptors. The crystal structures of SHP2 reveal an allosteric regulation of its activity. Under basal conditions, the N-SH2 domain blocks the catalytic site of the PTP domain. Association of SHP2 to its binding partners through the SH2 domains favors the release of this autoinhibitory interaction, making the catalytic site available to substrates. Although it is evident that the binding of a phosphopeptide on the N-SH2 domain promotes its displacement from PTP, the details of the molecular mechanisms, underlying the SHP2 activation or the allostery of the N-SH2 domain, was unclear and largely debated at the time of the proposal. For that reason, the purpose of this project was to use a combination of computational methods, X-ray scattering techniques, and biochemical experiments to identify at the atomic level the activation mechanism that brings to the opening of SHP2 in solution and, in addition, to build an atomistic model that explains how the disease-associated mutations perturb such a mechanism. The achievement of those objectives is an essential condition for the design of new molecules able to interfere specifically with the interaction of the SHP2 mutants to their partners in signal transduction and modulating its function.
Data: CORDIS, © European Union
Project objective
SHP2 is an SH2 domain-containing protein tyrosine phosphatase with a key role in the RAS-MAPK signaling pathway. Germline mutations in PTPN11, the gene encoding SHP2, occur in 50% of individuals affected by Noonan syndrome, whereas somatic mutations in this gene cause more than 30% of cases of juvenile myelomonocytic leukemia (JMML), and are more rarely found in other hematologic malignancies and tumors.The X-ray structure of SHP2 shows a multidomain architecture compatible with an allosteric regulatory mechanism: under basal conditions SHP2 is inactive, because its N-terminal Src homology 2 (N-SH2) domain blocks the active site of the protein tyrosine phosphatase (PTP) domain. Binding of the N-SH2 domain to a phosphopeptide (PP) ligand causes SHP2 activation by favoring dissociation of the N-SH2 and PTP domains.To characterize this dynamic transition at the atomic level, will be used a combination of state of the art computational methods coupled to X-ray scattering (XS) and biochemical assays. Major goals of the proposed studies are to explain how disease-associated mutations perturb the regulatory events controlling SHP2 function, and design new molecules able to inhibit SHP2 binding to signaling partners.The project implementation is guaranteed by the expertise of the Experienced Researcher (ER) in molecular dynamics (MD) studies of allosteric mechanisms that is complemented by the consolidated expertise of the host institution (HI) in the combined use of MD and XS, together with the external collaboration of an Italian expert in SHP2 biology and a US specialist in XS. Overall, the proposed research will provide a deeper understanding of SHP2 regulation, and dissect the molecular mechanisms implicated in its functional dysregulation in human disease. The planned studies are expected to provide a new class of lead compounds to treat SHP2-associated diseases. Finally, a new computational approach for the design of peptidomimetic inhibitors will be developed.
Original text from CORDIS.
Participants
- GEORG-AUGUST-UNIVERSITAT GOTTINGEN STIFTUNG OFFENTLICHEN RECHTS · GottingenCoordinatorGermany
Links
Data: CORDIS, © European Union