Crystallization and preliminary diffraction analysis of the T2I.L262F double mutant form of SHP2

Abstract

The enzymatic activity of SHP2, whose dysregulation causes malfunctions in diverse cellular signaling, is controlled by the autoinhibitory association between its N-SH2 and phosphatase domains. Various SHP2 genetic mutations, which impair the interaction between the two domains, have been identified to cause RAS-MAPK pathway-associated diseases. In this study, SHP2 containing Noonan syndrome-associated T2I and L262F double mutations was targeted for crystallization. The recombinant protein was prepared using an Escherichia coli expression system, purified using Ni-NTA affinity and size exclusion chromatographies, and then subjected for crystallization. X-ray data diffracted to 3.0 A resolution were collected and used for preliminary diffraction analysis. The unit cell parameters of the crystals belonging to the P21 space group were a = 45.4 A, b = 215.0 A, c = 55.6 A, and β = 95.7°. The asymmetric unit contains two molecules with a solvent content of 40.7% and a Matthews coefficient of 2.07 A3/Da.