Purification and characterization of a human urine ribonuclease (RNAase 1) showing genetic polymorphism.
Purification and characterization of a human urine ribonuclease (RNAase 1) showing genetic polymorphism.
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显示遗传多态性的人尿核糖核酸酶 (RNAase 1) 的纯化和表征。
DOI：
10.1016/0304-4165(88)90055-4
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
K. Kishi
中科院分区：
文献类型：
--
作者：
Toshihiro Yasuda;Wataru Sato;K. Kishi
通过 DEAE-Sepharose CL-6b、磷酸纤维素和 CM-纤维素柱层析，然后在 Bio-Gel P-100 和 Sephadex G-75 上进行凝胶过滤，最后通过 SDS-聚丙烯酰胺凝胶电泳达到均质状态，从 45 岁男性的尿液中分离纯化出核糖核酸酶 (RNAase)。该酶被命名为 RNAase 1。使用等电聚焦电泳，然后用特定于 RNAase 1 的兔抗体进行免疫印迹，可以轻松检测尿液和血清中的 RNAase 1 同工酶。利用该技术检测人血清中是否存在 RNAase 1 遗传多态性（Yasuda, T. 等人（1988）Am. J. Hum. Gent.，出版中）。就分子量和pI值而言，血清和尿液中的RNA酶1似乎以多种形式存在。遗传多态性RNA酶1是一种糖蛋白，每个分子含有3个甘露糖、1个岩藻糖、4个葡萄糖胺，不含唾液酸残基，凝胶过滤和SDS-聚丙烯酰胺凝胶电泳测定其分子量分别为16 000和17 500。该酶在 pH 7.0 时对酵母 RNA 底物最活跃，并且受到 Cu2+、Hg2+ 和 Zn2+ 的显着抑制。它还显示出对聚（C）和聚（U）的明确底物偏好，但对聚（A）和聚（G）的活性要低得多。因此，该酶是嘧啶特异性RNA酶。
A ribonuclease (RNAase) was isolated and purified from the urine of a 45-year-old man by column chromatographies on DEAE-Sepharose CL-6b, cellulose phosphate and CM-cellulose followed by gel filtrations on Bio-Gel P-100 and Sephadex G-75, and finally to a homogeneous state by SDS-polyacrylamide gel electrophoresis. The enzyme was designated RNAase 1. It was possible to detect RNAase 1 isozymes in urine and serum without difficulty using isoelectric focusing electrophoresis followed by immunoblotting with a rabbit antibody specific to RNAase 1. The existence of genetic polymorphism of RNAase 1 was detected in human serum utilizing this technique (Yasuda, T. et al. (1988) Am. J. Hum. Gent., in press). RNAase 1 in serum and urine seemed to exist in multiple forms with regard to molecular weight andpI value. Genetically polymorphic RNAase 1 was a glycoprotein, containing three mannose, one fucose, four glucosamine and no sialic acid residues per molcule, with a molecular weight of 16 000 and 17 500 determined by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively. The enzyme was most active at pH 7.0 on yeast RNA substrate adn inhibited remarkably by Cu2+, Hg2+and Zn2+. It also showed definite substrate preference for poly(C) and poly(U), but much less activity against poly(A) and poly(G). Thus, the enzyme is a pyrimidine-specific RNAase.
影响因子：
2.9
作者：
BLANK, A;DEKKER, CA
通讯作者：
DEKKER, CA