An electrophoretic separation and a quantitative staining method for leucine aminopeptidase, arylamidase and cystyl aminopeptidase were described. L-Leucinamide was used as a substrate for the aminopeptidases electrophoretically separated, and the liberated L-leucine was oxidized by leucine dehydrogenase and NAD+. The activity bands were visualized by tetrazolium salt (MTT) as a final hydrogen acceptor. Arylamidase and cystyl aminopeptidase were stained in the area of pre α1-and between α1 andα2-globulin position, respectively. The leucine aminopeptidase was samely stained in the area of preβ-globulin. Employing this method, it was possible to determine the activity of each aminopeptidase with satisfactory accuracy and reproducibility. Furthermore, this method would be useful to the analysis of enzymological properties of aminopeptidases because of its availability to other peptides, such as L-leucylglycine and L-leucyl-L-leucine, as substrate.
Leucine Dehydrogenaseを共役させたヒトAminopeptidaseの定量的活性染色法
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