Quantitation of autoinhibitory defects in pathogenic SHP2 mutants by differential scanning fluorimetry.

Src-homology-2-domain-containing protein tyrosine phosphatase-2 (SHP2) is a signaling enzyme whose activity is governed by an equilibrium between autoinhibited and activated states. Regulation of SHP2 activity is critical for cellular homeostasis, and mutations that alter its autoregulatory equilibrium cause cancers and developmental disorders. Several methods for assessing the strength of autoinhibitory interactions in SHP2 mutants have been previously reported, but each has limitations. We show that differential scanning fluorimetry provides a rapid, quantitative measure of SHP2 autoinhibition that is independent of the intrinsic activity of the SHP2 mutant being analyzed, does not involve protein labeling, and does not require specialized instrumentation. [Display omitted] • SHP2's activity is controlled by an interdomain autoinhibitory interaction. • Mutations that weaken SHP2's autoinhibition are causative for human pathologies. • Assessing SHP2 autoinhibition generally requires specialized equipment. • Differential scanning fluorimetry provides a rapid measure of SHP2 autoinhibition. • DSF does not require protein labeling or specialized instrumentation. [ABSTRACT FROM AUTHOR]