Stinson R A, Seargeant L E
Clin Chim Acta. 1981 Mar 5;110(2-3):261-72. doi: 10.1016/0009-8981(81)90355-7.
The alkaline phosphatases present in the human tissues liver, kidney, intestine, placenta and a serum from a patient with Paget's disease of bone have been purified to apparent homogeneity by affinity elution from a phosphonic acid-Sepharose derivative. Polyacrylamide gel electrophoresis in SDS gave subunit molecular weights ranging from 74 000 for the enzyme form placenta to 95 800 for the enzyme from kidney. The purified native and desialylated enzymes have been characterized by agar electrophoresis and isoelectric focusing. All five of the native enzymes behaved differently but the desialylated forms from liver, Paget's serum and kidney were indistinguishable in both systems. The desialylated enzyme from placenta and the enzyme from intestine behaved differently from each other and from the above mentioned group. The isoelectric points ranged from less than 4 for the native enzyme from liver to 7.01 for the desialylated forms of the enzymes from liver, kidney and Paget's serum. The effects of a number of factors on the thermostability of the purified enzymes were studied. Phosphate decreased the stability and human serum albumin increased the stability of most molecular forms. Desialylation had no effect on thermostability. These results and those from inhibition studies with L-phenylalanine, L-homoarginine, phosphate and vanadate support the 3-gene hypothesis advanced from structural studies.
通过从膦酸 - 琼脂糖衍生物上进行亲和洗脱，已将存在于人体组织肝脏、肾脏、肠道、胎盘以及一名患有骨佩吉特病患者血清中的碱性磷酸酶纯化至表观均一。在SDS中的聚丙烯酰胺凝胶电泳显示，亚基分子量范围从胎盘来源的酶的74000到肾脏来源的酶的95800。已通过琼脂电泳和等电聚焦对纯化的天然酶和去唾液酸酶进行了表征。所有五种天然酶表现不同，但肝脏、佩吉特病血清和肾脏来源的去唾液酸形式在这两种系统中无法区分。胎盘来源的去唾液酸酶和肠道来源的酶彼此之间以及与上述组的行为不同。等电点范围从肝脏来源的天然酶的小于4到肝脏、肾脏和佩吉特病血清来源的酶的去唾液酸形式的7.01。研究了多种因素对纯化酶热稳定性的影响。磷酸盐降低稳定性，而人血清白蛋白增加大多数分子形式的稳定性。去唾液酸化对热稳定性没有影响。这些结果以及用L - 苯丙氨酸、L - 高精氨酸、磷酸盐和钒酸盐进行的抑制研究结果支持了从结构研究提出的三基因假说。
