The type 5, acid phosphatase from spleen of humans with hairy cell leukemia. Purification, properties, immunological characterization, and comparison with porcine uteroferrin.
The type 5, acid phosphatase from spleen of humans with hairy cell leukemia. Purification, properties, immunological characterization, and comparison with porcine uteroferrin.
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DOI：
10.1016/s0021-9258(18)89088-3
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发表时间：
1985-05
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影响因子：
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通讯作者：
Catherine;Ketcham;George;BaurnbachSQll;Fuller;Bazers;R. Roberts
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作者：
Catherine;Ketcham;George;BaurnbachSQll;Fuller;Bazers;R. Roberts
毛细胞白血病患者的脾脏含有高水平的酒石酸盐不敏感的阳离子酸性磷酸酶（人类5型同工酶）。这种磷酸酶已被纯化的程序，其中只涉及两个色谱步骤：CM-纤维素层析和免疫亲和层析对猪子宫铁蛋白产生的羊抗体。子宫铁蛋白是一种丰富的含铁酸性磷酸酶，可以很容易地从猪子宫分泌物中回收。与子宫铁蛋白一样，纯化的人5型磷酸酶是一种分子量约为34，000的糖蛋白。它含有两个铁原子/分子。人磷酸酶和子宫铁蛋白在电泳迁移率、底物特异性和对多种激活剂和抑制剂的反应方面也彼此密切相似。已经产生了针对子宫铁蛋白和针对人5型磷酸酶的小鼠单克隆抗体。以高亲和力结合子宫铁蛋白分子上不同位点的三种单克隆抗体也识别人脾酶，但以低得多的亲和力结合。这些抗体还识别从牛和大鼠脾脏纯化的阳离子酸性磷酸酶。针对人类酶的单克隆抗体，但选择结合子宫铁蛋白，似乎认识到所有四个磷酸酶相对保守的网站。我们的结论是，人类5型同工酶属于一类结构相关的，含铁的酸性磷酸酶，其中包括铁转运蛋白，子宫铁蛋白。
The spleens of patients with hairy cell leukemia contain high levels of a tartrate-insensitive, cationic, acid phosphatase (the human Type 5 isozyme). This phosphatase has been purified by a procedure which involves only two chromatographic steps: CM-cellulose chromatography and immunoaffinity chromatography on sheep antibodies generated against porcine uteroferrin. Uteroferrin is an abundant iron-containing acid phosphatase that can be recovered readily from porcine uterine secretions. Like uteroferrin, the purified human Type 5 phosphatase is a glycoprotein of molecular weight about 34,000. It contains two atoms of iron/molecule. The human phosphatase and uteroferrin also resemble each other closely in electrophoretic mobility, substrate specificity, and response to a variety of activators and inhibitors. Mouse monoclonal antibodies have been raised to uteroferrin and to the human Type 5 phosphatase. Three monoclonal antibodies which bind with high affinities to distinct sites on the uteroferrin molecule also recognize the human spleen enzyme, but bind to it with much lower affinity. These antibodies also recognize cationic acid phosphatases purified from bovine and rat spleens. A monoclonal antibody raised against the human enzyme, but selected for binding to uteroferrin, appears to recognize a relatively conserved site on all four phosphatases. We conclude that the human Type 5 isozyme belongs to a growing class of structurally related, iron-containing acid phosphatases which includes the iron-transport protein, uteroferrin.