Fig. 5
From: Auto-thiophosphorylation activity of Src tyrosine kinase
Activation of Src by auto-thiophosphorylation. a Unphosphorylated Src catalytic domain (2.2Ā Ī¼M) was preincubated for 15Ā min at 30Ā Ā°C either alone, in the presence of 2Ā mM ATP and 10Ā mM MgCl2, or in the presence of 2Ā mM ATPĪ³S and 10Ā mM NiCl2. Src activity was measured using the phosphocellulose paper assay in the presence of peptide (0.8Ā mM) and [32P]-ATP (0.4Ā mM). Error bars show standard deviations. b Unphosphorylated Src (1Ā Ī¼M) was preincubated for various lengths of time at 30Ā Ā°C with 2Ā mM ATPĪ³S and 10Ā mM NiCl2. Src activity was measured toward the synthetic peptide substrate (0.87Ā mM) using the continuous spectrophotometric assay