NADH-coupled spectrophotometric assay of tyrosine aminotransferase.
A rapid spectrophotometric method for estimation of tyrosine aminotransferase activity (TAT) is described, based on a coupled reaction with NADH-dependent aromatic ketoacid reductase. 3-iodo-L-tyrosine, upon TAT action, is transformed into 3-iodo-4-hydroxyphenylpyruvate which quickly reacts with NADH in the presence of aromatic ketoacid reductase; oxidation rates at 340 nm are linear with protein concentration over the whole range of purification steps of TAT. This new method, for its sensitivity, easy performance and possibility of a continuous monitoring of TAT reaction, may be considered comparable to the more diffuse spectrophotometric standard method, and also as an alternative, advantageous procedure in some instances. The method for purification of the coupled aromatic ketoacid reductase is also described.
D000429:Alcohol Oxidoreductases; D000818:Animals; D006863:Hydrogen-Ion Concentration; D008297:Male; D007470:Monoiodotyrosine; D009243:NAD; D009994:Osmolar Concentration; D010667:Phenylpyruvic Acids; D011817:Rabbits; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014444:Tyrosine Transaminase / Q000032:analysis*
D010667:Phenylpyruvic Acids; D009243:NAD; D000429:Alcohol Oxidoreductases; C020060:diiodophenylpyruvate reductase; D014444:Tyrosine Transaminase; D007470:Monoiodotyrosine