生化分析揭示了 RCAN1/Rcn1 对钙调素抑制作用的新见解。

Yan Ren, Hui Chen, Shan-Yue Zhao, Lei Ma, Qing-Xia He, Wei-Bin Gong, Jia-Wei Wu, Hong-Wei Yao, Zhi-Xin Wang
{"title":"生化分析揭示了 RCAN1/Rcn1 对钙调素抑制作用的新见解。","authors":"Yan Ren,&nbsp;Hui Chen,&nbsp;Shan-Yue Zhao,&nbsp;Lei Ma,&nbsp;Qing-Xia He,&nbsp;Wei-Bin Gong,&nbsp;Jia-Wei Wu,&nbsp;Hong-Wei Yao,&nbsp;Zhi-Xin Wang","doi":"10.1111/febs.17266","DOIUrl":null,"url":null,"abstract":"<p>Calcineurin is a serine/threonine protein phosphatase that is highly conserved from yeast to human and plays a critical role in many physiological processes. Regulators of calcineurin (RCANs) are a family of endogenous calcineurin regulators, which are capable of inhibiting the catalytic activity of calcineurin <i>in vivo</i> and <i>in vitro</i>. In this study, we first characterized the biochemical properties of yeast calcineurin and its endogenous regulator Rcn1, a yeast homolog of RCAN1. Our data show that Rcn1 inhibits yeast calcineurin toward pNPP substrate with a noncompetitive mode; and Rcn1 binds cooperatively to yeast calcineurin through multiple low-affinity interactions at several docking regions. Next, we reinvestigated the mechanism underlying the inhibition of mammalian calcineurin by RCAN1 using a combination of biochemical, biophysical, and computational methods. In contrast to previous observations, RCAN1 noncompetitively inhibits calcineurin phosphatase activity toward both pNPP and phospho-RII peptide substrates by targeting the enzyme active site in part. Re-analysis of previously reported kinetic data reveals that the RCAN1 concentrations used were too low to distinguish between the inhibition mechanisms [Chan B <i>et al.</i> (2005) <i>Proc Natl Acad Sci USA</i> 102, 13075]. The results presented in this study provide new insights into the interaction between calcineurin and RCAN1/Rcn1.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Biochemical analyses reveal new insights into RCAN1/Rcn1 inhibition of calcineurin\",\"authors\":\"Yan Ren,&nbsp;Hui Chen,&nbsp;Shan-Yue Zhao,&nbsp;Lei Ma,&nbsp;Qing-Xia He,&nbsp;Wei-Bin Gong,&nbsp;Jia-Wei Wu,&nbsp;Hong-Wei Yao,&nbsp;Zhi-Xin Wang\",\"doi\":\"10.1111/febs.17266\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Calcineurin is a serine/threonine protein phosphatase that is highly conserved from yeast to human and plays a critical role in many physiological processes. Regulators of calcineurin (RCANs) are a family of endogenous calcineurin regulators, which are capable of inhibiting the catalytic activity of calcineurin <i>in vivo</i> and <i>in vitro</i>. In this study, we first characterized the biochemical properties of yeast calcineurin and its endogenous regulator Rcn1, a yeast homolog of RCAN1. Our data show that Rcn1 inhibits yeast calcineurin toward pNPP substrate with a noncompetitive mode; and Rcn1 binds cooperatively to yeast calcineurin through multiple low-affinity interactions at several docking regions. Next, we reinvestigated the mechanism underlying the inhibition of mammalian calcineurin by RCAN1 using a combination of biochemical, biophysical, and computational methods. In contrast to previous observations, RCAN1 noncompetitively inhibits calcineurin phosphatase activity toward both pNPP and phospho-RII peptide substrates by targeting the enzyme active site in part. Re-analysis of previously reported kinetic data reveals that the RCAN1 concentrations used were too low to distinguish between the inhibition mechanisms [Chan B <i>et al.</i> (2005) <i>Proc Natl Acad Sci USA</i> 102, 13075]. The results presented in this study provide new insights into the interaction between calcineurin and RCAN1/Rcn1.</p>\",\"PeriodicalId\":94226,\"journal\":{\"name\":\"The FEBS journal\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-09-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The FEBS journal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/febs.17266\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The FEBS journal","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/febs.17266","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

钙调素是一种丝氨酸/苏氨酸蛋白磷酸酶,从酵母到人类都高度保守,在许多生理过程中发挥着关键作用。钙调素调节剂(RCANs)是内源性钙调素调节剂的一个家族,能够抑制体内和体外钙调素的催化活性。在这项研究中,我们首先描述了酵母钙调素及其内源调节因子 Rcn1(RCAN1 的酵母同源物)的生化特性。我们的数据显示,Rcn1 以非竞争方式抑制酵母钙调素对 pNPP 底物的作用;Rcn1 通过多个对接区域的多重低亲和力相互作用与酵母钙调素合作结合。接下来,我们结合生化、生物物理和计算方法,重新研究了 RCAN1 抑制哺乳动物钙调蛋白的机制。与之前的观察结果不同,RCAN1 通过部分靶向酶活性位点,非竞争性地抑制了钙调磷酸酶对 pNPP 和磷酸化 RII 肽底物的活性。对以前报道的动力学数据的重新分析表明,使用的 RCAN1 浓度太低,无法区分抑制机制[Chan B 等人 (2005) Proc Natl Acad Sci USA 102, 13075]。本研究的结果为了解钙调蛋白与 RCAN1/Rcn1 之间的相互作用提供了新的视角。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Biochemical analyses reveal new insights into RCAN1/Rcn1 inhibition of calcineurin

Biochemical analyses reveal new insights into RCAN1/Rcn1 inhibition of calcineurin

Calcineurin is a serine/threonine protein phosphatase that is highly conserved from yeast to human and plays a critical role in many physiological processes. Regulators of calcineurin (RCANs) are a family of endogenous calcineurin regulators, which are capable of inhibiting the catalytic activity of calcineurin in vivo and in vitro. In this study, we first characterized the biochemical properties of yeast calcineurin and its endogenous regulator Rcn1, a yeast homolog of RCAN1. Our data show that Rcn1 inhibits yeast calcineurin toward pNPP substrate with a noncompetitive mode; and Rcn1 binds cooperatively to yeast calcineurin through multiple low-affinity interactions at several docking regions. Next, we reinvestigated the mechanism underlying the inhibition of mammalian calcineurin by RCAN1 using a combination of biochemical, biophysical, and computational methods. In contrast to previous observations, RCAN1 noncompetitively inhibits calcineurin phosphatase activity toward both pNPP and phospho-RII peptide substrates by targeting the enzyme active site in part. Re-analysis of previously reported kinetic data reveals that the RCAN1 concentrations used were too low to distinguish between the inhibition mechanisms [Chan B et al. (2005) Proc Natl Acad Sci USA 102, 13075]. The results presented in this study provide new insights into the interaction between calcineurin and RCAN1/Rcn1.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信