SPRING通过取代抑制性前结构域许可S1P介导的SREBP2切割-动脉网

SPRING licenses S1P-mediated cleavage of SREBP2 by displacing an inhibitory pro-domain

Nature 等信源发布 2024-07-09 08:34

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AbstractSite-one protease (S1P) conducts the first of two cleavage events in the Golgi to activate Sterol regulatory element binding proteins (SREBPs) and upregulate lipogenic transcription. S1P is also required for a wide array of additional signaling pathways. A zymogen serine protease, S1P matures through autoproteolysis of two pro-domains, with one cleavage event in the endoplasmic reticulum (ER) and the other in the Golgi.

AbstractSite one蛋白酶（S1P）在高尔基体中进行两次切割事件中的第一次，以激活固醇调节元件结合蛋白（SREBPs）并上调脂肪生成转录。S1P也是多种其他信号通路所必需的。S1P是一种酶原丝氨酸蛋白酶，通过两个pro结构域的自蛋白水解而成熟，一个在内质网（ER）中发生裂解事件，另一个在高尔基体中发生裂解事件。

We recently identified the SREBP regulating gene, (SPRING), which enhances S1P maturation and is necessary for SREBP signaling. Here, we report the cryo-EM structures of S1P and S1P-SPRING at sub-2.5 Å resolution. SPRING activates S1P by dislodging its inhibitory pro-domain and stabilizing intra-domain contacts.

我们最近鉴定了SREBP调节基因（SPRING），它增强了S1P的成熟，并且是SREBP信号传导所必需的。。SPRING通过去除其抑制性前结构域并稳定结构域内接触来激活S1P。

Functionally, SPRING licenses S1P to cleave its cognate substrate, SREBP2. Our findings reveal an activation mechanism for S1P and provide insights into how spatial control of S1P activity underpins cholesterol homeostasis..

在功能上，SPRING允许S1P切割其同源底物SREBP2。我们的发现揭示了S1P的激活机制，并为S1P活性的空间控制如何支持胆固醇稳态提供了见解。。

IntroductionSite-one protease (S1P, also known as Membrane-bound transcription factor site-1 protease (MBTPS1), or Subtilisin kexin isozyme 1 (SKI-1)), is a membrane-bound protease that mediates the proteolytic activation of transcription factors, hormones, and enzymes required for lipogenesis, the endoplasmic reticulum (ER) stress response, and lysosomal biogenesis, among others1,2.

引言site one蛋白酶（S1P，也称为膜结合转录因子site-1蛋白酶（MBTPS1）或枯草杆菌蛋白酶kexin同工酶1（SKI-1））是一种膜结合蛋白酶，可介导转录因子，激素的蛋白水解激活。脂肪生成，内质网（ER）应激反应和溶酶体生物发生等所需的酶1,2。

The best-studied role for S1P is in cholesterol homeostasis, where it conducts the first of two cleavage steps that mature the transcription factors Sterol regulatory element binding proteins (SREBPs) to upregulate cholesterol uptake and biosynthesis3,4. Genetic deletion of S1P is embryonically lethal in mice5 and zebrafish6, and the deletion of S1P in mouse livers results in decreased cholesterol and fatty acid biosynthesis and decreased plasma cholesterol5.

S1P研究得最好的作用是胆固醇稳态，它进行两个裂解步骤中的第一个，成熟转录因子固醇调节元件结合蛋白（SREBPs）以上调胆固醇摄取和生物合成3,4。S1P的遗传缺失在小鼠5和斑马鱼6中具有胚胎致死性，小鼠肝脏中S1P的缺失导致胆固醇和脂肪酸生物合成减少，血浆胆固醇降低5。

In humans, hypomorphic variants in MBTPS1 have been linked to skeletal dysplasia and elevated lysosomal enzymes in the blood7,8. Additionally, S1P is exploited by Arena viruses to mature their viral glycoproteins9,10,11 and by Hepatitis C virus (HCV), where S1P and SREBP are required to regulate the viral lifecycle12.

在人类中，MBTPS1的亚型变异与骨骼发育异常和血液中溶酶体酶升高有关7,8。此外，S1P被Arena病毒利用以成熟其病毒糖蛋白9,10,11和丙型肝炎病毒（HCV），其中S1P和SREBP需要调节病毒生命周期12。

Based on mutational analysis, the S1P cleavage motif is often described as RXXK↓ or RXXL ↓ . Mutation of the P4 Arg invariably eliminates cleavage by S1P and mutating the P1 residue to Ala also inhibits proteolysis of certain S1P substrates13,14,15. A more complete description of the motif is also reported as RX(L/I/V)Z, where X is residues other than Pro and Cys, and Z is preferably Lys or Leu and is not Val, Pro, Cys, or Glu1.

基于突变分析，S1P切割基序通常被描述为RXXK↓或RXXL↓。P4 Arg的突变总是消除S1P的切割，并且将P1残基突变为Ala也抑制某些S1P底物的蛋白水解13,14,15。基序的更完整描述也被报道为RX（L/I/V）Z，其中X是Pro和Cys以外的残基，Z优选Lys或Leu，而不是Val，Pro，Cys或Glu1。

S1P substrate preferences are distinct among the subtilisin-like proteases of the mammalian secretory pathway and the structural basis for the specificity of S1P activity is not known.Beginning fr.

S1P底物偏好在哺乳动物分泌途径的枯草杆菌蛋白酶样蛋白酶中是不同的，S1P活性特异性的结构基础尚不清楚。开始fr。

Data availability

数据可用性

The atomic models generated in this study are deposited in the Protein Data Bank (PDB) with codes PDB 8UW8 (S1Pecto-SPRINGecto) and PDB 8UWC(S1Pecto) and the cryo-EM maps are deposited in the Electron Microscopy Data Bank (EMDB) with the accession codes: EMD-42639 (S1Pecto-SPRINGecto) and EMD-42661 (S1Pecto).

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The raw movies have been deposited to EMPIAR with the accession codes: EMPIAR-11928 (S1Pecto-SPRINGecto) and EMPIAR-11931 (S1Pecto). Atomic models generated previously that were used in this study are PDB 4Z2A and PDB 2PMW. Source data are provided with this paper. All other data are available from the corresponding authors upon request. Source data are provided with this paper..

原始电影已存入EMPIAR，登记号为：EMPIAR-11928（S1Pecto SPRINGecto）和EMPIAR-11931（S1Pecto）。先前在本研究中使用的原子模型是PDB 4Z2A和PDB 2PMW。本文提供了源数据。所有其他数据可应要求从通讯作者处获得。本文提供了源数据。。

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Download referencesAcknowledgementsThis project was supported in part by R00GM141261 (to DLK). HH was supported through the Dr. Emmett J. Conrad Leadership Program. N.Z. is an Established Investigator of the Dutch Heart Foundation (2013T111) and is supported by a Vici grant from the Netherlands Organization for Scientific Research (NWO; 016.176.643) and an NWO ENW grant (M.22.034; GENESIS).

下载参考文献致谢R00GM141261（向DLK）部分支持该项目。HH得到了Emmett J.Conrad博士领导力计划的支持。N、 Z.是荷兰心脏基金会（2013T111）的资深研究员，并得到荷兰科学研究组织（NWO；016.176.643）和NWO ENW资助（M.22.034；GENESIS）的Vici资助。

All cryo-EM data was collected at the UT Southwestern Cryo-Electron Microscopy Facility (CEMF). We thank Dan Stoddard, Ph.D., and the CEMF staff for assistance with cryo-EM data collection. The CEMF is supported by a core facilities award from the Cancer Prevention & Research Institute of Texas (CPRIT RP220582).

。我们感谢Dan Stoddard博士和CEMF工作人员在低温电磁数据收集方面的帮助。CEMF得到了德克萨斯州癌症预防与研究所（CPRIT RP220582）颁发的核心设施奖的支持。

We thank Chad Brautigam and Shih-Chia Tso for carrying out SEC-MALS and Mass Photometry experiments. We thank members of the Zelcer and Kober labs and Irith Koster for their critical comments and suggestions on this study.Author informationAuthors and AffiliationsDepartment of Medical Biochemistry, Amsterdam UMC, Amsterdam Cardiovascular Sciences and Gastroenterology and Metabolism, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, the NetherlandsSebastian Hendrix, Jenina Kingma & Noam ZelcerDepartment of Biochemistry, The University of Texas Southwestern Medical Center, Dallas, TX, 75390, USAVincent Dartigue, Hailee Hall, Shrankhla Bawaria, Bilkish Bajaj & Daniel L.

我们感谢Chad Brautigam和Shih Chia Tso进行SEC-MALS和质量光度测定实验。我们感谢Zelcer和Kober实验室的成员以及Irith Koster对这项研究的批判性评论和建议。作者信息作者和附属机构阿姆斯特丹大学医学生物化学系，阿姆斯特丹UMC，阿姆斯特丹心血管科学与胃肠学与代谢系，阿姆斯特丹大学，Meibergdreef 91105AZ，阿姆斯特丹，荷兰巴斯蒂安·亨德里克斯，Jenina Kingma＆Noam Zelcer德克萨斯大学西南医学中心生物化学系，德克萨斯州达拉斯，75390，USAVENT Dartigue，Hailee Hall，Shrankhla Bawaria，Bilkish Bajaj＆Daniel L。

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PubMed Google ScholarContributionsS.H. conceptualized the study, developed and carried out western blot experiments and revised and edited the paper. V.D. developed the protease assay, determined the enzyme kinetics, and developed and carried out the competitive coIP assay. H.H.

PubMed谷歌学术贡献。H、将研究概念化，开发并进行了蛋白质印迹实验，并修订和编辑了论文。五、。H、 H。

carried out the protein purifications for cryo-EM grid preps, carried out biochemical characterization of the protein samples, and conducted initial coIP assays. S.B. carried out protease assays. J.K. carried out western blot experiments. B.B. carried out with molecular biology and protein expression experiments.

进行了冷冻EM网格制备的蛋白质纯化，对蛋白质样品进行了生化表征，并进行了初步的coIP分析。S、 B.进行蛋白酶测定。J、 K.进行了蛋白质印迹实验。B、 B.通过分子生物学和蛋白质表达实验进行。

N.Z. conceived the project, supervised research, and wrote the paper. D.L.K. conceived the project, supervised research, did the cryo-EM analysis, carried out biochemical assays, and wrote the paper.Corresponding authorsCorrespondence to.

N、 Z.构思了这个项目，监督了研究，并撰写了论文。D、 L.K.构思了该项目，监督了研究，进行了低温电磁分析，进行了生化分析，并撰写了论文。通讯作者通讯。

Noam Zelcer or Daniel L. Kober.Ethics declarations

诺姆·泽尔或丹尼尔·科伯。道德宣言

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Reprints and permissionsAbout this articleCite this articleHendrix, S., Dartigue, V., Hall, H. et al. SPRING licenses S1P-mediated cleavage of SREBP2 by displacing an inhibitory pro-domain.

转载和许可本文引用本文Hendrix，S.，Dartigue，V.，Hall，H。等人。SPRING通过置换抑制性前结构域来许可S1P介导的SREBP2切割。

Nat Commun 15, 5732 (2024). https://doi.org/10.1038/s41467-024-50068-8Download citationReceived: 17 February 2024Accepted: 28 June 2024Published: 09 July 2024DOI: https://doi.org/10.1038/s41467-024-50068-8Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

《国家公社》155732（2024）。https://doi.org/10.1038/s41467-024-50068-8Download引文接收日期：2024年2月17日接受日期：2024年6月28日发布日期：2024年7月9日OI：https://doi.org/10.1038/s41467-024-50068-8Share本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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