Isolation and characterizaion of renin processing enzyme from renin granule

肾素颗粒中肾素加工酶的分离和表征

基本信息

批准号：
04670567
负责人：
IKEDA Masaharu
金额：
$ 1.28万
依托单位：
University of Occupational & Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670567/
关键词：
Renin Prorenin Processing Renin Granule Subtilisin プレセシング

项目摘要

Processing mechanism of prorenin to renin is still controversial. It is postulated that renin is synthesized and processed in the rough endoplasmic reticulum and the Golgi complex and stored in the renin granules and secreted by exocytosis. Recently several processing enzymes which process prohormones or protein precursors were found and identified as intrinsic processing enzymes.The purpose of this study was to clarify the nature of the intrinsic renin processing enzyme in the subcellular fraction of the kidney cortex. Generally prohormone convertases or protein precursor processing enzymes were reported to a common catalytic domain homologous to that of the subtilisin serine protease family.Hence in the first step of the present study, we investigated whether subtilisin per se can process prorenin to renin. When radiolabelled[^<35>S]-human recombinant prorenin was incubated with subtilisin BNP^-, human recombinant prorenin which showed a molecular weight of 43,000 dalton was cleaved … More to renin, a molecular weight of 38,000 dalton. In addition, when semipurified human amniotic prorenin(or inactive renin) was incubated with subtilisin, we confirmed the activation of prorenin by subtilisin by detection of active renin which was quantitated by a specific immunoradiometric assay system. These results indicate that subtilisin per se can process and activate prorenin to active form.In the next study, we investigated whether renin granule fraction of the kidney can process recombinant human prorenin to renin by cleaving prosegment of the prorenin. Human prorenin (molecular weight 43,000) was cleaved to renin (molecular weight 38,000) by renin granule fraction. This result suggests that renin granule fraction contained prorenin processing enzyme. In this study, we tried to isolate the prorenin processing enzyme from renin granule fraction, but we could noto isolate or identified prorenin processing enzyme in the renin granule because of loss of activity probably due to its lability during isolation. Less

prorenin到肾素的加工机制仍然存在争议。据推测，肾素是在粗糙的内质网和高尔基体配合物中合成和处理的，并储存在肾素颗粒中，并被胞吞作用分泌。最近，发现了几种加工性激素或蛋白质前体的加工酶，并将其鉴定为内在加工酶。本研究的目的是阐明肾脏皮质亚细胞分数中固有的肾素加工酶的性质。通常，据报道了与枯草酶蛋白SE蛋白酶家族同源的常见催化结构域的蛋白转化酶或蛋白质前体加工酶。因此，在本研究的第一步中，我们研究了枯草脂本质上是否可以在肾素上加工前肾蛋白。当放射性标记的[^<35> s] - 人类重组prorenin与枯草蛋白酶bnp^ - 孵育时，人类重组prorenin显示出43,000 dalton的分子量，裂解了43,000个dalton…更多地到肾素，一种分子量，一种分子量，分子量为38,000 Dalton。此外，当将半纯化的人羊膜释蛋白（或非活性肾素）与枯草酶一起孵育时，我们通过检测活性肾蛋白来证实prorenin通过枯草素的激活，该肾素通过特定的免疫放射测定系统定量。这些结果表明，枯草素本身可以处理并激活prorenin至活性形式。在下一项研究中，我们研究了肾脏的肾素颗粒分数是否可以通过切割prose摄影来处理重组人的prorenin，从而将肾素处理为肾素。肾素颗粒馏分将人prorenin（分子量43,000）裂解至肾素（分子量38,000）。该结果表明，肾素颗粒分数含有prorenin加工酶。在这项研究中，我们试图从肾素颗粒的分数中分离出prorenin的加工酶，但是由于活性损失可能是由于其在分离过程中的衰退，因此无法分离或鉴定肾素颗粒中的prorenin加工酶。较少的