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Intracellular Biophotonic Nanoswitches

细胞内生物光子纳米开关

基本信息

批准号：
EP/F040954/1
负责人：
Rudolf Allemann
金额：
$ 183.55万
依托单位：
CARDIFF UNIVERSITY
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2008
资助国家：
英国
起止时间：
2008 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FF040954%2F1
关键词：
Intracellular Biophotonic Nanoswitches

项目摘要

Recent mapping of all physical interactions between proteins in a given cell has confirmed the notion that interactions between proteins are highly regulated and underpin all cellular processes. Researchers and technologists have been presented with a major challenge - how to ask specific questions of such complex systems especially when protein interactions change with time in a given cell and result in different end states. For example when a human cell responds to stress, specific interactions between master regulatory proteins start to drive a recovery process or initiate a controlled commitment to cell death. This project aims to generate a generic technology for solving this problem - introducing synthetic switches into live cells that can 'fine-tune' protein interactions by remote control. This exciting approach, based on highly promising preliminary work, would allow the investigator to programme changes in defined protein-protein interactions by the introduction of small interfering molecules engineered to be switched on and off by light of carefully selected wavelengths. Changes in the structure of a small molecule are triggered by external light pulses inducing conformational rearrangements in the peptide backbone and hence alterations of the biological properties of the Intracellular Biophotonic Nanoswitch (IBN). IBNs are light-sensitive nanoparticle-based molecular structures linked to the short peptide sequences that recognize features on the surface of a protein that has been targeted for switching. Conventional and novel methods for IBN delivery into live cells will allow patterning of the swiches into populations of cells. Operating these IBNs by light will allow the researcher to pattern the activation of switches in such complex cell populations or to 'programme' the switching process in single cells - a step-forward in the technology of manipulating master regulators of discrete intracellular pathways. Our proposal's adventure and risk relates to the problems of IBN design and their potential for self-reporting in live cells.IBNs will allow a researcher to switch or programme the state of a master regulator in a live cell by biophysical means and explore the consequences on the whole system to reveal the internal linking of different pathways. Furthermore, our proposal addresses how to track the downstream consequences of selective switching, even in different lineages, to reveal how cells respond to different signals (amplitude or frequency) in developing their responses even if these arise quickly or indeed develop slowly through different cell generations.Since our vision is to provide the life sciences community with novel, robust and readily implemented technologies based on robust chemical systems, the proposal encompasses engagement with potential user & downstream demands of IBN technology with a focus on the burgeoning demand to understand cellular biology at the complex 'systems' level. The exciting prospect looms of gaining programmable photonic control over normal physiology (directing stem cell differentiation, manipulating wound healing and delaying cell senescence), neoplasia (cancer biology of cell cycle checkpoint dysfunction and photonically-controlled therapeutics), constructed cell communities (light-directed tissue engineering) and molecular target identification (the search for new medicines and products).

最近对给定细胞中蛋白质之间的所有物理相互作用的绘图证实了蛋白质之间的相互作用受到高度调节并支撑所有细胞过程的概念。研究人员和技术人员面临着一个重大挑战-如何提出这种复杂系统的具体问题，特别是当给定细胞中蛋白质相互作用随时间变化并导致不同的最终状态时。例如，当人类细胞对压力做出反应时，主调节蛋白之间的特定相互作用开始驱动恢复过程或启动受控的细胞死亡。该项目旨在产生一种解决这一问题的通用技术-将合成开关引入活细胞，可以通过远程控制“微调”蛋白质相互作用。这种令人兴奋的方法，基于非常有前途的初步工作，将允许研究人员通过引入小的干扰分子来改变定义的蛋白质-蛋白质相互作用，这些干扰分子被精心选择的波长的光打开和关闭。小分子结构的变化由外部光脉冲触发，诱导肽骨架中的构象重排，从而改变细胞内生物光子纳米开关（IBN）的生物学性质。IBN是基于光敏纳米颗粒的分子结构，其连接到短肽序列，该短肽序列识别已被靶向转换的蛋白质表面上的特征。用于IBN递送到活细胞中的常规和新颖方法将允许将交换物图案化成细胞群。通过光操作这些IBN将使研究人员能够在这种复杂的细胞群中激活开关，或者“编程”单细胞中的开关过程-这是操纵离散细胞内途径的主调节器技术的一个进步。我们的建议的冒险和风险涉及IBN设计的问题和它们在活细胞中自我报告的潜力。IBN将允许研究人员通过生物物理手段切换或编程活细胞中主调节器的状态，并探索对整个系统的影响，以揭示不同途径的内部联系。此外，我们的建议涉及如何跟踪选择性转换的下游后果，即使在不同的谱系中，以揭示细胞如何响应不同的信号（幅度或频率）在发展它们的反应中起作用，即使这些反应在不同的细胞代中迅速出现或确实缓慢发展。基于强大的化学系统的强大和易于实施的技术，该提案包括与IBN技术的潜在用户和下游需求的接触，重点是在复杂的“系统”水平上了解细胞生物学的新兴需求。令人兴奋的前景隐约可见，即获得对正常生理学（指导干细胞分化，操纵伤口愈合和延迟细胞衰老），肿瘤（细胞周期检查点功能障碍的癌症生物学和光子控制疗法），构建细胞群落（光导向组织工程）和分子靶点识别（寻找新药物和产品）的可编程光子控制。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

¹H, ¹³C and ¹5N chemical shift assignments of unliganded Bcl-xL and its complex with a photoresponsive Bak-derived peptide.

未配体的 Bcl-xL 及其与光响应 Bak 衍生肽的复合物的 H、C 和 5N 化学位移分配。

DOI：
10.1007/s12104-012-9407-9
发表时间：
2013
期刊：
Biomolecular NMR assignments
影响因子：
0.9
作者：
Wysoczanski P
通讯作者：
Wysoczanski P

Real-time cell viability assays using a new anthracycline derivative DRAQ7®.

DOI：
10.1002/cyto.a.22228
发表时间：
2013-02
期刊：
CYTOMETRY PART A
影响因子：
3.7
作者：
Akagi, Jin;Kordon, Magdalena;Zhao, Hong;Matuszek, Anna;Dobrucki, Jurek;Errington, Rachel;Smith, Paul J.;Takeda, Kazuo;Darzynkiewicz, Zbigniew;Wlodkowic, Donald
通讯作者：
Wlodkowic, Donald

Chemical synthesis of cell-permeable apoptotic peptides from in vivo produced proteins.

DOI：
10.1021/bc200338u
发表时间：
2011-08
期刊：
Bioconjugate chemistry
影响因子：
4.7
作者：
T. Fricke;R. J. Mart;C. Watkins;M. Wiltshire;R. Errington;Paul J. Smith;A. Jones;R. Allemann
通讯作者：
T. Fricke;R. J. Mart;C. Watkins;M. Wiltshire;R. Errington;Paul J. Smith;A. Jones;R. Allemann

Photocontrolled Exposure of Pro-apoptotic Peptide Sequences in LOV Proteins Modulates Bcl-2 Family Interactions.