Landemore G, Buat M L, Izard J
Biol Cell. 1987;59(1):97-100. doi: 10.1111/j.1768-322x.1987.tb00518.x.
Following previous light and ultrastructural cytochemical reports, this paper presents the first zymograms of Kurloff cell acid phosphatases extracted from highly purified cell suspensions. Using isoelectric focusing, up to 20 isoenzymes were observed both with hexazotised pararosanilin or fast garnet GBC as coupler and naphthol ASB1 phosphate as substrate. Most were tartrate-labile. After Clostridium-derived neuraminidase digestion, the highly stained bands observed at pH 4-5 disappeared and, simultaneously, a few alkaline bands were enhanced. Two main bands of Mr 190,000 and 500,000 were characterized by their acid phosphatase activity after electrophoresis on a 4-15% gradient native polyacrylamide gel.
继之前的光镜和超微结构细胞化学报告之后,本文展示了从高度纯化的细胞悬液中提取的库尔洛夫细胞酸性磷酸酶的首张酶谱图。使用等电聚焦法,以重氮对玫瑰苯胺或固红GBC作为偶联剂、萘酚ASB1磷酸酯作为底物时,观察到多达20种同工酶。大多数对酒石酸不稳定。经梭菌来源的神经氨酸酶消化后,在pH 4 - 5处观察到的高染色带消失,同时,一些碱性带增强。在4 - 15%梯度的天然聚丙烯酰胺凝胶上进行电泳后,通过其酸性磷酸酶活性鉴定出两条主要条带,分子量分别为190,000和500,000。