Sugiyama R H, Blank A, Dekker C A
Biochemistry. 1981 Apr 14;20(8):2268-74. doi: 10.1021/bi00511a031.
Four major urine ribonuclease (RNase) activities, designated bands A-D, were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and activity staining. Bands A, B, and C have alkaline pH optima and display molecular weights of 31 000, 23 000, and 20 000, respectively, upon sodium dodecyl sulfate (NaDodSO4) gel electrophoresis and weights of 44 000, 28 000, 22 000 upon gel filtration. Band D, with a pH optimum slightly below neutrality, has a molecular weight of 16 000 or 15 000, respectively, determined by the above methods. Band A, the most abundant activity in urine, is heterogeneous and resembles serum RNase 1 on electrophoresis and on phosphocellulose and Sephadex chromatography. Band B is similar to a minor, unnamed component of serum RNase activity while band C resembles serum RNase 3. Band D is similar to the leukocyte RNase-like activity of serum [Blank, A., & Dekker, C.A. (1981) Biochemistry (preceding paper in this issue)]. Band A is present in urine at a concentration high than that of RNase 1 in serum. In contrast, urine counterparts of serum RNases 2, 4, and 5 are not apparent upon either phosphocellulose chromatography [see also Yamanaka, M., Akagi, K., Murai, K., Hirao, N., Fujimi, S., & Omae, T. (1977) Clin. Chim. Acta 78, 191-201] or NaDodSO4 get electrophoresis; a urine counterpart of serum RNase 3 can be detected only by the more sensitive electrophoretic method. These results indicate that RNase 2-5 are processed differently by the kidney than RNase 1. After reconciliation of reported differences in their pH optima and molecular weights, five apparently diverse RNase preparations described in the literature can be related to band A activity and three preparations to band D. However, we are unable to confirm a previous report of a human urine enzyme indistinguishable from bovine pancreatic RNase A.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和活性染色，鉴定出四种主要的尿核糖核酸酶（RNase）活性，分别命名为A-D条带。A、B和C条带的最适pH呈碱性，在十二烷基硫酸钠（NaDodSO4）凝胶电泳中，其分子量分别为31000、23000和20000，而在凝胶过滤中分子量分别为44000、28000、22000。D条带的最适pH略低于中性，通过上述方法测定其分子量分别为16000或15000。A条带是尿液中最丰富的活性成分，具有异质性，在电泳以及磷酸纤维素和葡聚糖凝胶色谱上类似于血清RNase 1。B条带类似于血清RNase活性中的一种次要的、未命名的成分，而C条带类似于血清RNase 3。D条带类似于血清中的白细胞RNase样活性[布兰克，A.，& 德克尔，C.A.（1981年）《生物化学》（本期之前的论文）]。A条带在尿液中的浓度高于血清中RNase 1的浓度。相比之下，无论是通过磷酸纤维素色谱法[另见山中，M.，赤木，K.，村井，K.，平尾，N.，藤见，S.，& 大前，T.（1977年）《临床化学学报》78，191 - 201]还是十二烷基硫酸钠凝胶电泳，血清RNase 2、4和5在尿液中的对应物都不明显；血清RNase 3的尿液对应物只能通过更灵敏的电泳方法检测到。这些结果表明，肾脏对RNase 2 - 5的处理方式与RNase 1不同。在协调了它们报道的最适pH和分子量差异后，文献中描述的五种明显不同的RNase制剂可与A条带活性相关，三种制剂与D条带相关。然而，我们无法证实先前关于一种与牛胰RNase A无法区分的人尿酶的报道。
