RUI: Understanding Cu(I) Interactions with Zinc-Metalloproteins: Impact of Cu(I) on Structure and Function of the X-Linked Inhibitor of Apoptosis Protein

RUI：了解 Cu(I) 与锌金属蛋白的相互作用：Cu(I) 对 X 连锁凋亡蛋白抑制剂结构和功能的影响

• 批准号: 1807773
• 负责人: Kathryn Splan
• 金额: $ 29.27万
• 依托单位: Macalester College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1807773&HistoricalAwards=false
• 关键词: RUI; Understanding; Cu; Interactions; Zinc

This award from the Chemistry of Life Processes Program to Profesor Kathryn Splan of Macalester College supports research that investigates the interaction of copper ions with zinc-binding proteins. Many proteins require metal ions for proper function, and replacement of the usual metal ion with one exhibiting different chemical properties is likely to alter protein structure, and therefore, function. While metal-ion substitution has clear ramifications in the context of metal-ion toxicity, changes in protein function upon metal-ion substitution may also constitute a desirable functional strategy. The experiments reveal the susceptibility of zinc proteins toward substitution by copper and characterize the impact of this substitution on the protein's function. The project trains students in biochemical preparation and characterization techniques, thereby building solid technical skills in preparation for further study and/or entry into the work force. The results from the project also inform the fields of cell biology, neurochemistry, and neurobiology, particularly in those areas concerned with the molecular basis of diseases that are characterized by mis-regulated metal ion levels.In this work, the impact of Cu(I) on metalloprotein function is further investigated by characterizing the interaction of Cu(I) with the X-linked Inhibitor of Apoptosis Protein (XIAP), a Zn(II)-metalloprotein recently implicated in copper homeostasis. Multiple Zn(II) protein structural motifs have been shown to be susceptible to displacement by Cu(I), owing to the shared thiophilic character of both metal ions. Cu(I) substitution at the Zn(II) structural sites of XIAP is one possible mechanism by which XIAP function may be modulated by changing copper concentrations. A variety of spectroscopic and biochemical approaches are utilized to detect the displacement of zinc ions upon the addition of copper and the resulting structural and functional changes. Understanding Cu(I) interactions with this target at the molecular level deepens the understanding of metalloprotein susceptibility toward disruption by Cu(I) in general and clarifies the role of Cu(I) in XIAP function.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.

这项由生命过程化学项目授予麦卡莱斯特学院的凯瑟琳·斯普兰的奖项支持了研究铜离子与锌结合蛋白相互作用的研究。许多蛋白质需要金属离子才能发挥正常功能，用具有不同化学性质的金属离子取代通常的金属离子可能会改变蛋白质的结构，从而改变功能。虽然金属离子取代在金属离子毒性的背景下具有明确的分支，但金属离子取代后蛋白质功能的变化也可能构成理想的功能策略。实验揭示了锌蛋白对铜取代的敏感性，并表征了这种取代对蛋白质功能的影响。该项目培训学生生化制备和表征技术，从而建立坚实的技术技能，为进一步学习和/或进入劳动力市场做准备。该项目的结果也为细胞生物学、神经化学和神经生物学领域提供了信息，特别是在那些与以金属离子水平失调为特征的疾病的分子基础有关的领域。在这项工作中，通过表征Cu（I）与X-连锁凋亡抑制蛋白（XIAP）的相互作用，一种最近与铜稳态有关的锌（II）金属蛋白。多个Zn（II）蛋白质结构基序已被证明易受Cu（I）置换的影响，这是由于两种金属离子共享的亲硫特性。在XIAP的Zn（II）结构位点处的Cu（I）取代是一种可能的机制，通过该机制，XIAP功能可以通过改变铜浓度来调节。利用各种光谱和生物化学方法来检测添加铜后锌离子的位移以及由此产生的结构和功能变化。理解Cu（I）在分子水平上与该目标的相互作用加深了对Cu（I）破坏的金属蛋白敏感性的理解，并阐明了Cu（I）在XIAP功能中的作用。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

项目成果

Disruption of zinc (II) binding and dimeric protein structure of the XIAP-RING domain by copper (I) ions

• DOI: 10.1007/s00775-023-02002-4
• 发表时间: 2023-06
• 期刊: JBIC Journal of Biological Inorganic Chemistry
• 作者: Kathryn E. Splan;Sylvia R Choi;Ruth E Claycomb;Isaiah K. Eckart-Frank;Shreya Nagdev;Madeline E Rodemeier

Emerging investigator series: characterization of silver and silver nanoparticle interactions with zinc finger peptides

• DOI: 10.1039/c9en00065h
• 发表时间: 2019-08-01
• 期刊: ENVIRONMENTAL SCIENCE-NANO
• 影响因子: 7.3
• 作者: Park, Grace;Amaris, Zoe N.;Wheeler, Korin E.
• 通讯作者: Wheeler, Korin E.