Paessens A, Rüdiger H
Eur J Biochem. 1980 Nov;112(1):47-51. doi: 10.1111/j.1432-1033.1980.tb04985.x.
The transferase was isolated by means of hydrophobic chromatography and combination of ion-exchange and gel filtration at different pH values and ionic strengths. As judged by disc electrophoresis, the enzyme is homogeneous. Electrophoresis in the presence of sodium dodecylsulfate reveals only one band with Mr = 49500 +/- 10%. In gel filtration the native enzyme has a Mr of 200,000. The subunits can be crosslinked by iminothiolane followed by hydrogen peroxide oxidation. In sodium dodecylsulfate electrophoresis this results in a band pattern of integer multiples of 50,000 up to 20,000 but not higher. The high-Mr aggregates disappear on splitting the crosslinks by reduction. Thus the enzyme appears to be composed of four subunits identical or nearly identical in Mr. By the dansyl method, only phenylalanine and methionine were found as the amino-terminal residues, suggesting the existence of two different types of subunits.
通过疏水色谱法以及在不同pH值和离子强度下结合离子交换和凝胶过滤的方法分离出了转移酶。通过圆盘电泳判断，该酶是均一的。在十二烷基硫酸钠存在下进行电泳，仅显示出一条Mr = 49500±10%的条带。在凝胶过滤中，天然酶的Mr为200,000。亚基可以通过亚胺硫醇烷交联，然后用过氧化氢氧化。在十二烷基硫酸钠电泳中，这导致出现高达20,000但不高于此值的50,000的整数倍的条带模式。通过还原断裂交联键，高Mr聚集体消失。因此，该酶似乎由四个Mr相同或几乎相同的亚基组成。通过丹磺酰法，仅发现苯丙氨酸和甲硫氨酸作为氨基末端残基，这表明存在两种不同类型的亚基。
