Acquisition of a Confocal Fluorescence Microscope to Image Synthetic Biomolecular Condensates

获取共焦荧光显微镜对合成生物分子凝聚物进行成像

基本信息

批准号：
10581200
负责人：
Ashutosh Chilkoti
金额：
$ 19.79万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10581200
关键词：
Administrative Supplement Alanine Area Behavior Biochemical Biocompatible Materials Biotechnology Books Cells Cholesterol Confocal Microscopy Core Facility Crowding Development Diffuse Disease Elastin Environment Eukaryotic Cell Evolution Exhibits Face Fluorescence Microscopy Fluorescence Recovery After Photobleaching Funding Genes Grant Image In Vitro Investigation Kinetics Length Libraries Measures Medicine Methods Microscope Modification Molecular Morphology National Institute of General Medical Sciences Oligonucleotides Parents Peptides Phase Phase Transition Phosphorylation Polymers Post-Translational Protein Processing Property Proteins Recombinants Research Research Support Reservations Scanning Self-Direction Signal Transduction Stimulus Structural Protein Structure System Temperature Tertiary Protein Structure Time Work aqueous cell behavior design equipment acquisition fluorescence microscope genetic information heuristics in vivo insight instrument interest microsystems myristoylation parent grant polypeptide recombinant peptide response self assembly spatiotemporal

项目摘要

Abstract The proposed work in this MIRA application leverages my long-standing interest and expertise in the design of genetically encoded stimulus-responsive peptide polymers. My group pioneered the development of recombinant elastin-like polypeptides (ELPs) that exhibit lower critical solution temperature (LCST) phase behavior. We have also, in parallel, pioneered the development of high-throughput methods for the assembly of highly repetitive genes that we used to create the largest extant library of recombinant peptide polymers. Characterization of their aqueous phase behavior led to the discovery of sequence heuristics that can be used for the de novo used design of peptide polymers that exhibit LCST phase behavior and a class of resilin-like polypeptides (RLPs) that exhibit the converse — upper critical solution phase transition (UCST) phase behavior. Building upon this work, we will explore two new areas in this proposal. First, we will investigate how we can recapitulate the hierarchical structure and properties exhibited by biological materials by the design of partially ordered polymers (POPs) —that consist of disordered polypeptides embedded with a periodically recurring secondary structure motif— that exhibit temperature triggered hierarchical self-assembly into macroscopic materials that mimic the in vivo organization of structural proteins like elastin networks. We will carry out a systematic exploration of the design of new POPs, to verify that the combination of order and disorder at the chain segment level is a new and robust design principle that will yield materials with hierarchical selfassembly across many length scales. Second, we will develop a new line of investigation on genetically encoded biohybrid polymers via post-translational modifications (PTMs) that precisely combine peptide and non-peptide components to create biomaterials that exhibit triggered, hierarchical self-assembly into macroscopic materials. In this aim, we will expand upon our initial work on in vivo myristoylation of ELPs and UCST exhibiting RLPs to investigate if we can convert structure-directing peptides into myristoylation substrates, to create myristoylated polypeptides where the myristoylated segment can direct hierarchical self- assembly of the entire construct. We will also investigate modification of ELPs and RLPs with cholesterol that has the potential to direct self-assembly, and phosphorylation, which will provide a unique trigger of self- assembly. Much remains to be done in both areas, as our preliminary foray into these new areas only hint at the enormous possibilities in the molecular design of new biomaterials enabled by these approaches. The work we propose herein promises to yield new biomaterials with interesting structures and properties with a host of applications in biotechnology and medicine.

抽象的此MIRA应用中的拟议工作利用了我在设计方面的长期兴趣和专业知识遗传编码的刺激反应性肽聚合物。我的小组开创了发展暴露于较低临界溶液温度（LCST）相的重组弹性蛋白样多肽（ELP）行为。我们还同时还开创了用于组装的高通量方法的开发我们用来创建最大现存的重组肽聚合物库的高度重复基因的基因。其水相行为的表征导致发现可以使用的序列启发式方法对于从头开始使用的辣椒设计，暴露了LCST阶段行为和一类类似于溶质蛋白暴露了相反的多肽（RLP） - 上临界溶液相变（UCST）相行为。在这项工作的基础上，我们将探索该提案中的两个新领域。首先，我们将调查如何我们可以通过设计通过生物材料来概括生物学材料的层次结构和特性部分有序的聚合物（POPS） - 由无序的多肽组成反复出现的二级结构图案 - 暴露温度触发层次自组装到模仿弹性蛋白等结构蛋白的体内组织的宏观材料。我们将对新流行音乐的设计进行系统的探索，以验证订单和连锁部门级别的障碍是一种新的稳固的设计原则，它将产生材料在许多长度尺度上进行分层自组装。其次，我们将开发一项新的调查。通过翻译后修饰（PTMS），一般编码的生物杂交聚合物，这些聚合物精确地结合了胡椒和非肽成分，以产生暴露于触发的生物材料，分层自组装进入宏观材料。在此目标中，我们将扩展我们关于ELPS肉豆蔻酰胺化的最初工作 UCST展示了RLP，以调查我们是否可以将结构导向肽转化为肉豆蔻酰化底物，创建肉豆蔻酰化的多肽，在该多肽中，肉豆蔻酰化段可以指导分层自我整个结构的组装。我们还将调查用胆固醇的ELP和RLP的修饰有可能指导自组装和磷酸化，这将提供自我的独特触发因素集会。在这两个领域仍有很多待在这些方法实现了新生物材料的分子设计中的巨大可能性。工作我们在此提出，有望产生具有有趣结构和特性的新生物材料生物技术和医学中的应用。

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Engineering synthetic biomolecular condensates.

DOI：
10.1038/s44222-023-00052-6
发表时间：
2023-04-17
期刊：
Nature reviews bioengineering
影响因子：
0
作者：
Dai, Yifan;You, Lingchong;Chilkoti, Ashutosh
通讯作者：
Chilkoti, Ashutosh

Genetically Engineered Nanoparticles of Asymmetric Triblock Polypeptide with a Platinum(IV) Cargo Outperforms a Platinum(II) Analog and Free Drug in a Murine Cancer Model.

DOI：
10.1021/acs.nanolett.2c01850
发表时间：
2022-07-27
期刊：
NANO LETTERS
影响因子：
10.8
作者：
Saha, Soumen;Banskota, Samagya;Liu, Jianqiao;Zakharov, Nikita;Dzuricky, Michael;Li, Xinghai;Fan, Ping;Deshpande, Sonal;Spasojevic, Ivan;Sharma, Kedar;Borgnia, Mario Juan;Schaal, Jeffrey L.;Raman, Ashutosh;Kim, Sarah;Bhattacharyya, Jayanta;Chilkoti, Ashutosh
通讯作者：
Chilkoti, Ashutosh

Author Correction: Injectable tissue integrating networks from recombinant polypeptides with tunable order.

作者更正：可注射组织整合来自具有可调顺序的重组多肽的网络。

DOI：
10.1038/s41563-018-0233-z
发表时间：
2018
期刊：
Nature materials
影响因子：
41.2
作者：
Roberts,Stefan;Harmon,TylerS;Schaal,JeffreyL;Miao,Vincent;Li,KanJonathan;Hunt,Andrew;Wen,Yi;Oas,TerrenceG;Collier,JoelH;Pappu,RohitV;Chilkoti,Ashutosh
通讯作者：
Chilkoti,Ashutosh

Genetically encoded lipid-polypeptide hybrid biomaterials that exhibit temperature-triggered hierarchical self-assembly.

DOI：
10.1038/s41557-018-0005-z
发表时间：
2018-05
期刊：
Nature chemistry
影响因子：
21.8
作者：
Mozhdehi D;Luginbuhl KM;Simon JR;Dzuricky M;Berger R;Varol HS;Huang FC;Buehne KL;Mayne NR;Weitzhandler I;Bonn M;Parekh SH;Chilkoti A
通讯作者：
Chilkoti A

Intrinsically disordered proteins access a range of hysteretic phase separation behaviors