Hammond K D, Wollbrandt R K, Gilbert D A
Int J Biochem. 1985;17(2):259-64. doi: 10.1016/0020-711x(85)90124-7.
The activities of acid and alkaline phosphatases and phosphotyrosine, phosphoserine and phosphothreonine phosphatases were measured in Friend murine erythroleukaemic (MEL) cells. The effects of treating the cells with dimethyl sulphoxide (DMSO), an inducer of differentiation, were examined. In untreated cells alkaline phosphatase activity was undetectable, though there were significant amounts of acid phosphatase (76 +/- 15 mU/mg protein) and phosphotyrosine phosphatase (16 +/- 0.9mU/mg protein); phosphoserine and phosphothreonine phosphatase activities (9 +/- 0.4 and 7 +/- 0.6mU/mg protein, respectively) were lower than for phosphotyrosine phosphatase. Addition of 1 or 2% DMSO to the culture medium resulted in the expected cell death within 2 weeks. With 0.5% DMSO, cells remained viable for at least 8 weeks, but while some appeared to have smaller nuclei and retained their rounded appearance, others became fibroblastic within several days and adhered to the culture vessel. The treated cells which had kept their morphology showed no difference in acid phosphatase activities as compared with untreated controls; phosphotyrosine phosphatase was lower (9 +/- 0.8mU/mg protein) and phosphoserine and phosphothreonine phophatases higher (11 +/- 0.5 and 10 +/- 0.4mU/mg protein, respectively) than in the controls. The Km values for p-nitrophenyl phosphate were similar in untreated and treated cells (0.069 and 0.068mM, respectively); for phosphotyrosine the Km value was lower in the treated cells (0.97mM) than in the controls (1.9mM).
在弗瑞德小鼠红白血病(MEL)细胞中测定了酸性和碱性磷酸酶以及磷酸酪氨酸、磷酸丝氨酸和磷酸苏氨酸磷酸酶的活性。研究了用二甲基亚砜(DMSO)(一种分化诱导剂)处理细胞的效果。在未处理的细胞中,碱性磷酸酶活性无法检测到,不过存在大量酸性磷酸酶(76±15 mU/mg蛋白质)和磷酸酪氨酸磷酸酶(16±0.9 mU/mg蛋白质);磷酸丝氨酸和磷酸苏氨酸磷酸酶活性(分别为9±0.4和7±0.6 mU/mg蛋白质)低于磷酸酪氨酸磷酸酶。向培养基中添加1%或2%的DMSO会在2周内导致预期的细胞死亡。添加0.5%的DMSO时,细胞至少存活8周,但一些细胞似乎细胞核较小且保持圆形外观,而另一些细胞在几天内变成成纤维细胞并附着在培养容器上。保持形态的处理细胞与未处理对照相比,酸性磷酸酶活性没有差异;磷酸酪氨酸磷酸酶较低(9±0.8 mU/mg蛋白质),磷酸丝氨酸和磷酸苏氨酸磷酸酶较高(分别为11±0.5和10±0.4 mU/mg蛋白质)。对硝基苯磷酸酯的Km值在未处理和处理细胞中相似(分别为0.069和0.068 mM);对于磷酸酪氨酸,处理细胞中的Km值(0.97 mM)低于对照(1.9 mM)。