CAREER: Quantification of Cellular Proteome Stress and Recovery Using Chemical Methods

职业：使用化学方法量化细胞蛋白质组压力和恢复

• 批准号: 1944973
• 负责人: Xin Zhang
• 金额: $ 74万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-15 至 2022-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1944973&HistoricalAwards=false
• 关键词: CAREER; Quantification; Cellular; Proteome; Stress

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Xin Zhang from Pennsylvania State University to develop new tools examining the effects of stresses on proteins in live cells, in real time. The proteins in cells must be maintained in a delicate balance between a folded state that is dissolved in water (soluble folded) and an solid state that does not dissolve in water (insoluble aggregated state). Excessive aggregation is bad for the cells but this phenomenon occurs when cells are exposed to stresses such as aging, heat and certain chemicals. To understand how protein stress relates to cellular function, Dr. Zhang focuses develops a new class of fluorescent sensors that light up when they come into contact with unfolded proteins in cells. This project provides a variety of new tools for real-time determinations of cellular protein stresses and recovery. The interdisciplinary nature of the program allows graduate and undergraduate students to gain training and expertise in modern chemical biology research. The project also includes an education plan called "Seeking Protein Aggregates Detectors". The central theme of this education plan is to integrate K-12 and undergraduate students in research and public engagement programs that discover new chemical detectors for protein aggregates from unexplored natural dyes and their interactions with egg white proteins. As existing proteome stress sensors are insufficient to enable quantitative studies on the dynamics of proteome stress and recovery, Dr. Zhang is developing a novel class of fluorogenic sensors, represented by the AgHalo sensor. This sensor uses changes in fluorescence intensity as a direct readout to quantify stress-induced protein aggregation. Building on this foundation, Dr. Zhang is developing a second fluorogenic proteome stress sensor, AgSNAP, which is orthogonal to AgHalo, thus enabling a set of two-color sensors. To detect protein refolding as a post-stress event, a fluorogenic proteome recovery sensor is being developed using anti-solvatochromic fluorophores. These sensors are used to quantify intracellular protein aggregation and refolding in cells that are subjected to chronic stresses. Mutant proteins that can form either soluble or insoluble aggregates are also targeted. Results of these experiments test the hypothesis that the soluble aggregates of mutant proteins cause a more severe proteome stress than the insoluble aggregates. These research goals enable a set of novel proteome stress and recovery sensors for the protein biochemistry community and provide quantitative knowledge of proteome aggregation and refolding in chronically stressed cells.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

凭借该奖项，化学部的生命过程化学项目正在资助宾夕法尼亚州立大学的张鑫博士开发新工具，实时检查压力对活细胞中蛋白质的影响。细胞内的蛋白质必须在溶解于水的折叠状态（可溶性折叠）和不溶解于水的固态（不溶性聚集状态）之间维持微妙的平衡。过度聚集对细胞不利，但当细胞暴露于老化、热和某些化学物质等压力时，就会发生这种现象。为了了解蛋白质应激与细胞功能的关系，张博士重点开发了一种新型荧光传感器，当它们与细胞中未折叠的蛋白质接触时会发光。 该项目提供了多种用于实时测定细胞蛋白质应激和恢复的新工具。 该计划的跨学科性质使研究生和本科生能够获得现代化学生物学研究的培训和专业知识。该项目还包括一项名为“寻求蛋白质聚集体检测器”的教育计划。 该教育计划的中心主题是将 K-12 学生和本科生纳入研究和公共参与项目，这些项目发现新的化学探测器，用于检测未开发的天然染料中的蛋白质聚集体及其与蛋清蛋白的相互作用。由于现有的蛋白质组应激传感器不足以定量研究蛋白质组应激和恢复的动态，张博士正在开发一类新型荧光传感器，以AgHalo传感器为代表。 该传感器使用荧光强度的变化作为直接读数来量化压力诱导的蛋白质聚集。在此基础上，张博士正在开发第二种荧光蛋白质组压力传感器AgSNAP，它与AgHalo正交，从而实现了一组双色传感器。为了检测应激后的蛋白质重折叠，正在开发一种使用抗溶剂化显色荧光团的荧光蛋白质组恢复传感器。这些传感器用于量化遭受慢性应激的细胞中的细胞内蛋白质聚集和重折叠。 可以形成可溶性或不溶性聚集体的突变蛋白也是目标。这些实验的结果检验了这样的假设：突变蛋白的可溶性聚集体比不溶性聚集体引起更严重的蛋白质组应激。 这些研究目标为蛋白质生物化学界提供了一套新型蛋白质组应激和恢复传感器，并提供了长期应激细胞中蛋白质组聚集和重折叠的定量知识。该奖项反映了 NSF 的法定使命，并通过使用基金会的评估进行评估，被认为值得支持。智力价值和更广泛的影响审查标准。

项目成果

Stress-induced protein disaggregation in the endoplasmic reticulum catalysed by BiP

BiP 催化内质网中应激诱导的蛋白质解聚

• DOI: 10.1038/s41467-022-30238-2
• 发表时间: 2022-05-06
• 期刊: Nature Communications
• 影响因子: 16.6
• 作者: Melo, Eduardo Pinho;Konno, Tasuku;Farace, Ilaria;Awadelkareem, Mosab Ali;Skov, Lise R.;Teodoro, Fernando;Sancho, Teresa P.;Paton, Adrienne W.;Paton, James C.;Fares, Matthew;Paulo, Pedro M. R.;Zhang, Xin;Avezov, Edward
• 通讯作者: Avezov, Edward

Four-dimensional nuclear speckle phase separation dynamics regulate proteostasis

四维核散斑相分离动力学调节蛋白质稳态

• DOI: 10.1126/sciadv.abl4150
• 发表时间: 2022-01-07
• 期刊: Science Advances
• 影响因子: 13.6
• 作者: Dion W;Ballance H;Lee J;Pan Y;Irfan S;Edwards C;Sun M;Zhang J;Zhang X;Liu S;Zhu B
• 通讯作者: Zhu B