Anreicherung der Leber-N-Acetyltransferase durch präparative Polyacrylamidgel-Elektrophorese und Bestimmung des Molekulargewichts /Purification of Human Liver Serotonin-/Isoniazid-N-Acetyltransferase by Preparative Polyacrylamidgel-Electrophoresis and Determination of Molecular Weight
Abstract Human liver N-acetyltransferase was purified by means of column chromatography on Sephadex G-100 (18 fold; yield: 60-70%) and preparative disc electrophoresis (40 fold; yield: 95 - 100%). A separation of the hypothetical “serotonin-N-acetyltransferase” and “INH-N-acetyltransferase” on the basis of a possible difference of the charge of the proteins could not be achieved (disc electrophoresis). Thus the assumption is confirmed that INH and Serotonin are substrates of one enzyme. The activity of N-acetyltransferase can be stabilized by adding thioglycolate, 2-mercaptoethanol, and cysteine. In some cases an activation of 100 to 200% could be observed. A molecular weight of about 26 500 was determined by the method of column chromatography on Sephadex as well as by calculating the sedimentation coefficient. Enzymes prepared by different procedures showed similar Kм-values.