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. 2020 Oct 31;11(3):750–762. doi: 10.1016/j.apsb.2020.10.021

Figure 4.

Figure 4

Biological characterization of his-tagged SHP2 recombinant proteins. (A) Purity and molecular weight analysis of his-tagged SHP2-WT (61.4 kDa), SHP2-PTP (35.0 kDa) and SHP2-E76A (61.4 kDa) proteins by Coomassie Brilliant Blue staining. (B) Basal enzyme activity comparison between SHP2 proteins in the absence of p-IRS1 peptide. 10 μmol/L DiFMUP was titrated with different concentration of protein ranging from 0.037 to 2.36 nmol/L. (C) Activated enzyme activity comparison between SHP2-WT and SHP2-PTP enzymes. 10 μmol/L DiFMUP and 0.5 nmol/L enzyme were titrated with different concentration of p-IRS1 peptide ranging from 0.02 to 10 μmol/L. (D) Enzyme kinetic parameters of DiFMUP dephosphorylation by SHP2-PTP (0.225 nmol/L), SHP2-E76A (0.15 nmol/L), and SHP2-WT (0.5 nmol/L) under basal (0 μmol/L p-IRS1 peptide) and fully activated (6 μmol/L p-IRS1 peptide) condition. Data are shown as mean ± SD; n ≥ 3.