Pösö H, Pegg A E
Biochim Biophys Acta. 1983 Sep 28;747(3):209-14. doi: 10.1016/0167-4838(83)90099-7.
Ornithine decarboxylase (EC 4.1.1.17) activity was about 3-times higher in Saccharomyces uvarum than in Saccharomyces cerevisiae in the middle of logarithmic growth. The enzyme from both sources was inactivated by alpha-difluoromethylornithine. When the binding of [5-14 C]difluoromethylornithine to yeast ornithine decarboxylase was studied it was shown that S. uvarum extracts contained about 40 ng of active ornithine decarboxylase per mg of cellular protein, and that of S. cerevisiae 10-12 ng of active enzyme. It appeared that S. uvarum ornithine decarboxylase could be highly purified by affinity chromatography, but S. cerevisiae enzyme did not bind to the same column. The purified preparation from S. uvarum had an Mr of 73 000 and a 100-fold purification (purified by conventional methods) of ornithine decarboxylase from S. cerevisiae had an Mr of 69 000 on a gel filtration column. When the purified S. ovarum ornithine decarboxylase was labelled with difluoromethylornithine, it co-eluted with native enzyme on a gel filtration column and it ran as a single band on polyacrylamide gel electrophoresis under denaturing conditions at a position corresponding to an Mr of 72 000, indicating that the active enzyme is a monomer. The loss of ornithine decarboxylase activity after addition of cycloheximide and spermidine to culture correlated with the decrease of the binding of difluoromethylornithine to protein.
在对数生长中期，葡萄汁酵母中的鸟氨酸脱羧酶（EC 4.1.1.17）活性比酿酒酵母中的高约3倍。来自这两种来源的酶都被α-二氟甲基鸟氨酸灭活。当研究[5-¹⁴C]二氟甲基鸟氨酸与酵母鸟氨酸脱羧酶的结合时，发现葡萄汁酵母提取物每毫克细胞蛋白中含有约40纳克活性鸟氨酸脱羧酶，而酿酒酵母中含有10 - 12纳克活性酶。似乎葡萄汁酵母的鸟氨酸脱羧酶可以通过亲和层析高度纯化，但酿酒酵母的酶不与同一柱子结合。从葡萄汁酵母中纯化得到的制剂的相对分子质量为73000，而用常规方法纯化的酿酒酵母鸟氨酸脱羧酶在凝胶过滤柱上的相对分子质量为69000。当用二氟甲基鸟氨酸标记纯化的葡萄汁酵母鸟氨酸脱羧酶时，它在凝胶过滤柱上与天然酶共洗脱，并且在变性条件下的聚丙烯酰胺凝胶电泳中作为一条单一的带迁移，其位置对应于相对分子质量72000，表明活性酶是单体。向培养物中添加环己酰亚胺和亚精胺后鸟氨酸脱羧酶活性的丧失与二氟甲基鸟氨酸与蛋白质结合的减少相关。
