RAISE: Design of co-assembling peptides as recombinant protein fusion tags for integrating enzymes into supramolecular hydrogels

RAISE：设计共组装肽作为重组蛋白融合标签，用于将酶整合到超分子水凝胶中

• 批准号: 1743432
• 负责人: Carol Hall
• 金额: $ 100万
• 依托单位: North Carolina State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1743432&HistoricalAwards=false
• 关键词: RAISE; Design; co; assembling; peptides

This RAISE project is jointly funded by the Biological and Environmental Interactions of Nanoscale Materials program in CBET Division in the Engineering Directorate, the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences in the Biological Sciences Directorate, and the Office of Integrative Activities. In this multi-disciplinary project, nanomaterial scaffolds, or hydrogels, with immobilized enzymes will be produced using proteins as a design template. The proposed method provides a novel alternative to conventional immobilization techniques which are more difficult and time consuming. This novel assembly technique is designed so that it selectively self-assembles into nano fiber structures. This targeted design and assembly of proteins into nanofibers is an exciting new research frontier that is largely unexplored with regard to both fundamental protein assembly and design of functional nanomaterials. A multi-disciplinary team of investigators will combine their expertise in modeling, imaging and characterization methods in order to demonstrate novel protein assembly and to advance fundamental understanding of protein assembly. The proposed technique will be an extraordinary improvement over the current state-of-the-art in protein assembly methods. Successful implementation of the proposed technique is anticipated to have a long-term broad impact on the engineering of novel functional materials for various biomedical and biotechnological applications. A variety of educational activities will be developed, including an iPad app on protein folding, and a video describing how computational methods can be used to design new biomaterials.The proposed method bypasses conventional immobilization techniques in which the nanostructured scaffold is functionalized post-assembly via reactions at its surface. Instead, enzymes are expressed by bacteria from recombinant DNA with peptide fusion tags that mediate their assembly into nanostructured materials. The key to the method is the use of fusion tags that co-assemble into fibrillar structures when mixed with a complementary peptide, but remain unassembled when pure. Co-assembling peptides are an exciting research frontier that is virtually unexplored in both protein biophysics and functional nanomaterials. The goals of the project are to demonstrate novel enzyme-functionalized supramolecular hydrogels, and to advance fundamental biophysical understanding necessary to predict peptide co-assembly. Three specific aims are proposed: (1) characterize the assembly process and resultant structure for known â-sheet nanofiber-forming co-assembling peptide pairs, (2) design and test new pairs of selectively co-assembling peptides and, (3) test co-assembling peptides as tags to immobilize proteins in nanostructured biomaterials. Each aim will leverage the computational modeling expertise of PI Hall, with expertise in spectroscopic and microscopic characterization of peptide nanofibers provided by Co-PI Hudalla and solid-state nuclear magnetic resonance expertise provide by Co-PI Paravastu. The proposed enzyme immobilization method will be an extraordinary improvement over the current state-of-the-art in protein immobilization methods and hence opens the door to addressing long-standing challenges in installing biologically-active, folded proteins into biomaterials. A variety of educational activities will be developed, including an iPad app on protein folding, and a video describing how computational methods can be used to design new biomaterials.

该RAISE项目由工程理事会CBET部门的纳米级材料的生物和环境相互作用计划，生物科学理事会分子和细胞生物科学部门的分子生物物理学计划以及综合活动办公室共同资助。在这个多学科项目中，将使用蛋白质作为设计模板来生产具有固定化酶的纳米材料支架或水凝胶。所提出的方法提供了一种新的替代传统的固定化技术，这是更困难和耗时。 这种新的组装技术被设计成选择性地自组装成纳米纤维结构。这种有针对性的设计和蛋白质组装成纳米纤维是一个令人兴奋的新的研究前沿，在基本蛋白质组装和功能性纳米材料的设计方面基本上是未开发的。一个多学科的研究团队将联合收割机结合他们在建模，成像和表征方法的专业知识，以证明新的蛋白质组装和推进蛋白质组装的基本理解。所提出的技术将是对目前蛋白质组装方法的一个非凡的改进。所提出的技术的成功实施预计将对各种生物医学和生物技术应用的新型功能材料的工程产生长期广泛的影响。将开发各种教育活动，包括一个关于蛋白质折叠的iPad应用程序，以及一个描述如何使用计算方法来设计新生物材料的视频。所提出的方法绕过了传统的固定技术，在传统的固定技术中，纳米结构支架通过其表面的反应在组装后功能化。相反，酶由细菌从具有肽融合标签的重组DNA表达，肽融合标签介导酶组装成纳米结构材料。该方法的关键是融合标签的使用，当与互补肽混合时，融合标签共组装成纤维状结构，但当纯时保持未组装。 共组装肽是一个令人兴奋的研究前沿，在蛋白质生物物理学和功能性纳米材料中几乎未被探索。该项目的目标是展示新型酶功能化超分子水凝胶，并推进预测肽共组装所需的基本生物物理理解。 提出了三个具体的目标：（1）表征已知的形成纳米纤维的共组装肽对的组装过程和所得结构，（2）设计和测试新的选择性共组装肽对，以及（3）测试共组装肽作为纳米结构生物材料中蛋白质的标签。每个目标都将利用PI Hall的计算建模专业知识，Co-PI Hudalla提供的肽纳米纤维的光谱和微观表征专业知识以及Co-PI Paravastu提供的固态核磁共振专业知识。所提出的酶固定化方法将是对蛋白质固定化方法的当前最新技术的非凡改进，因此为解决将生物活性折叠蛋白质安装到生物材料中的长期挑战打开了大门。 将开发各种教育活动，包括关于蛋白质折叠的iPad应用程序，以及描述如何使用计算方法设计新生物材料的视频。

项目成果

Characterising the throat diameter of through-pores in network structures using a percolation criterion

• DOI: 10.1080/00268976.2019.1654140
• 发表时间: 2019-08
• 期刊: Molecular Physics
• 影响因子: 1.7
• 作者: K. Wang;C. Hall

A multiscale coarse-grained model to predict the molecular architecture and drug transport properties of modified chitosan hydrogels

用于预测改性壳聚糖水凝胶的分子结构和药物转运特性的多尺度粗粒度模型

• DOI: 10.1039/d0sm01243b
• 发表时间: 2020
• 期刊: Soft Matter
• 影响因子: 3.4
• 作者: Singhal, Ankush;Schneible, John D.;Lilova, Radina L.;Hall, Carol K.;Menegatti, Stefano;Grafmüller, Andrea
• 通讯作者: Grafmüller, Andrea

On the liquid demixing of water + elastin-like polypeptide mixtures: bimodal re-entrant phase behaviour

关于水弹性蛋白样多肽混合物的液体分层：双峰重入相行为

• DOI: 10.1039/d0cp05013j
• 发表时间: 2021
• 期刊: Physical Chemistry Chemical Physics
• 影响因子: 3.3
• 作者: Lindeboom, Tom;Zhao, Binwu;Jackson, George;Hall, Carol K.;Galindo, Amparo
• 通讯作者: Galindo, Amparo

Computational Study of DNA-Cross-Linked Hydrogel Formation for Drug Delivery Applications

• DOI: 10.1021/acs.macromol.8b01505
• 发表时间: 2018-11
• 期刊: Macromolecules
• 影响因子: 5.5
• 作者: K. Wang;Tania Betancourt;C. Hall

Molecular complementarity and structural heterogeneity within co-assembled peptide β-sheet nanofibers

共组装肽β片纳米纤维内的分子互补性和结构异质性

• DOI: 10.1039/c9nr08725g
• 发表时间: 2020
• 期刊: Nanoscale
• 影响因子: 6.7
• 作者: Wong, Kong M.;Wang, Yiming;Seroski, Dillon T.;Larkin, Grant E.;Mehta, Anil K.;Hudalla, Gregory A.;Hall, Carol K.;Paravastu, Anant K.
• 通讯作者: Paravastu, Anant K.