A synthetic toolkit for the recombinant production of tyrosine phosphorylated proteins and peptides

用于重组生产酪氨酸磷酸化蛋白和肽的合成工具包

基本信息

批准号：
10673930
负责人：
Kristen M Naegle
金额：
$ 35.83万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10673930
关键词：
Acceleration Acidic Amino Acids Adoption Affinity Alternative Splicing Bar Codes Binding Sites Biochemical Biological Breast Cancer Cell C-terminal Charge Clinic Computer Models Darkness Development Engineering Ensure Epidermal Growth Factor Receptor Escherichia coli Goals Heart Human In Vitro LYN gene Libraries Malignant Neoplasms Mass Spectrum Analysis Measurement Measures Molecular Monitor Mutagenesis Pattern Penetration Peptides Phosphopeptides Phosphoproteins Phosphorylation Phosphorylation Site Phosphoserine Phosphothreonine Phosphotransferases Phosphotyrosine Physiological Post-Translational Protein Processing Process Production Proteins Proteome Reaction Reagent Recombinants Reproducibility Research Role SH3 Domains Signal Transduction Site Specificity System Tail Technology Testing Therapeutic Intervention Tyrosine Tyrosine Phosphorylation Work aggressive breast cancer anticancer research cancer therapy cost epithelial to mesenchymal transition flexibility improved interest invention malignant breast neoplasm phosphoproteomics polyproline protein folding protein function src-Family Kinases synthetic protein tool treatment response tumor progression

项目摘要

Project Summary Tyrosine phosphorylation (pTyr) is a post-translational modiﬁcation that can regulate protein function and interac- tions. Tyrosine phosphorylation often becomes dysregulated in cancer and therefore, understanding the effect of phosphorylation on protein function and within biochemical networks will be paramount to identifying therapeutic interventions in cancer. Unfortunately, much of tyrosine phosphorylation is referred to as the “dark phosphopro- teome”, because we either cannot detect it or we do not understand what its role is in regulating proteins and networks. The dark phosphoproteome exists as a result of limitations in tools for testing and measuring pTyr within proteins and networks. In previous work, we developed a synthetic toolkit that has superior yields of phys- iologically relevant tyrosine phosphorylation on proteins of interest, compared to existing approaches. The goal of this work is to: 1) harden this technology for increased reproducibility and ﬂexibility and 2) extend this technol- ogy to produce reagents, at a fraction of their current cost, for use in quantitative measurement and monitoring of phosphotyrosine sites in mass spectrometry-based workﬂows relevant to cancer progression and treatment response.

项目摘要酪氨酸磷酸化（PTYR）是一种翻译后修饰，可以调节蛋白质功能和间 tions。酪氨酸磷酸化通常在癌症中失调，因此了解蛋白质功能和生化网络内的磷酸化对于识别治疗至关重要癌症的干预措施。不幸的是，许多酪氨酸磷酸化被称为“深磷酸 teome”，因为我们要么无法检测到它，要么不了解其在调节蛋白中的作用以及网络。由于测试和测量PTYR的工具的局限性而存在深色磷蛋白酶在蛋白质和网络中。在以前的工作中，我们开发了一种合成工具包与现有方法相比，对感兴趣蛋白质的生物学相关酪氨酸磷酸化。目标这项工作是：1）加强这项技术以提高可重复性和灵活性，2）扩展此技术 - OGY以当前成本的一小部分生产试剂，用于定量测量和监测与癌症进展和治疗相关的基于质谱的工作流中的磷酸酪氨酸位点回复。