In vitro activity: TAME inhibits cyclin proteolysis in mitotic Xenopus egg extract with IC50 of 12 μM. TAME at concentration of 1-200 μM arrests interphase extract treated with recombinant cyclin B1/Cdc2 complex in mitosis, with stable cyclin B1 and phosphorylated Cdc27. TAME at concentration of 200 μM dramaticly inhibits the ubiquitin ligase activity of the Anaphase-Promoting Complex (APC), accompanied by reduced binding of Cdh1 to APC. TAME addition to interphase extract reduces Cdc20 association with the APC in a dose-dependent manner partly by binding directly to APC, and the contribution motif is the C-terminal isoleucine-arginine (IR) tail on APC. TAME is hydrolysed by trypsin with Km of 0.328 mM. TAME accelerates the ATP hydrolysis process about 12-fold. TAME interacts with β and γ phosphate and the adenine ring of ATP by the guanidinium group and the aromatic ring. TAME at concentration of 50 mM inhibits nutrient-induced germination and pressure-induced germination at 600 MPa in Bacillus subtilis. TAME induces a concentration dependent contractile response on ileal strips with EC50 of 4.3 x 103 M.
Kinase Assay: TAME at concentration of 1-200 μM arrests interphase extract treated with recombinant cyclin B1/Cdc2 complex in mitosis, with stable cyclin B1 and phosphorylated Cdc27. TAME at concentration of 200 μM dramaticly inhibits the ubiquitin ligase activity of the Anaphase-Promoting Complex (APC), accompanied by reduced binding of Cdh1 to APC. TAME addition to interphase extract reduces Cdc20 association with the APC in a dose-dependent manner partly by binding directly to APC, and the contribution motif is the C-terminal isoleucine-arginine (IR) tail on APC. TAME is hydrolysed by trypsin with Km of 0.328 mM. TAME accelerates the ATP hydrolysis process about 12-fold. TAME interacts with β and γ phosphate and the adenine ring of ATP by the guanidinium group and the aromatic ring. TAME at concentration of 50 mM inhibits nutrient-induced germination and pressure-induced germination at 600 MPa in Bacillus subtilis. TAME induces a concentration dependent contractile response on ileal strips with EC50 of 4.3 x 103 M.
Cell Assay: In mitotic Xenopus oocytes, TAME competes with the Cdc20 C-terminal IR-tail for APC binding to inhibit APC-dependent proteolysis. [3] TAME also stabilizes cyclin B1via terminating ubiquitination prenaturally. It slows the unmodified cyclin B1 initial ubiquitination. In the presence of TAME, ubiquitinated cyclin B1 is not able to promote Cdc20 binding to the APC. | 
