Rapid and large-scale purification and characterization of renin from mouse submaxillary gland.
Rapid and large-scale purification and characterization of renin from mouse submaxillary gland.
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小鼠颌下腺肾素的快速大规模纯化和表征。
DOI：
10.1016/0003-9861(82)90539-2
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发表时间：
1982
影响因子：
3.9
通讯作者：
Inagami,T
中科院分区：
文献类型：
--
作者：
Misono,KS;Holladay,LA;Murakami,K;Kuromizu,K;Inagami,T
对于肾素高度受限的底物特异性的结构基础知之甚少，其迄今为止已知的唯一功能是水解血管紧张素原中独特的亮基肽键，形成血管紧张素i。我们缺乏知识是因为我们无法大量纯化这种酶以进行结构研究。建立了一种快速、大规模纯化小鼠颌下腺肾素的两步柱层析方法。它可以以数百毫克的量分离酶，总收率为60%。通过聚丙烯酰胺凝胶电泳、等电聚焦和超离心研究得到的单条带表明产物的均匀性。通过在6mguanidine·HCl中的沉降平衡研究，肾素的分子量估计为36,000。沉积速度研究给出了一个单一的沉积边界，为2.58 × 10−13s。凝胶过滤得到的Stokes半径为27 Å。远紫外-圆二色光谱显示β-结构含量高（46%）。与肾肾素不同，颌下腺肾素不含氨基糖或中性糖。刀豆蛋白琼脂糖凝胶没有保留肾素活性。氨基酸分析、等电聚焦和氨基末端残基的测定结果表明，该蛋白与先前分离的肾素A相同，但数量少得多。
Little is known about the structural basis for the highly restricted substrate specificity of renin, whose only function known to date is to hydrolyze the unique leucyl peptide bond in the prohormone angiotensinogen to form angiotensin I. Our lack of knowledge is due to our inability to purify this enzyme in a large quantity sufficient for structural studies. A two-step column chromatographic method for rapid and large-scale purification of renin from mouse submaxillary gland has been developed. It allows isolation of the enzyme in a hundreds-of-milligrams quantity at an overall yield of 60%. Single bands obtained by polyacrylamide gel electrophoresis, isoelectric focusing, and the result of ultracentrifugal studies indicated homogeneity of the product. The moelcular weight of renin was estimated to be 36,000 by sedimentation equilibrium studies in 6mguanidine · HCl. Sedimentation velocity study gave a single sedimenting boundary with ans20,wof 2.58 × 10−13s. A Stokes radius of 27 Å was obtained by gel filtration. The far ultraviolet-circular dichroism spectrum indicated a high content of β-structure (46%). In contrast to renal renin, submaxillary gland renin does not contain amino sugars or neutral sugars. No renin activity was retained by concanavalin Aagarose gels. Results of amino acid analysis, isoelectric focusing, and determination of amino-terminal residues by the dansylchloride reaction, together indicated that this protein is identical with renin A isolated previously in a much smaller quantity.
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蛋白质化学的战略和策略。
影响因子：
4.1
作者：
B. Hartley
通讯作者：
B. Hartley
重氮酰基试剂对人肾素的抑制：该酶与其他蛋白酶的关系。
影响因子：
6.1
作者：
M. Mckown;R. I. Gregerman
通讯作者：
R. I. Gregerman
影响因子：
3.9
作者：
James C. Lee;S. N. Timasheff
通讯作者：
S. N. Timasheff
聚丙烯酰胺凝胶中蛋白质的等电聚焦。
DOI：
10.1016/0005-2795(72)90248-6
发表时间：
1972
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
O. Vesterberg
通讯作者：
O. Vesterberg
浓缩氯化胍中蛋白质溶液的沉降平衡。
影响因子：
2.9
作者：
P. Munk;D. Cox
通讯作者：
D. Cox