Photochemistry of Azide Based Cross-Linkers in Water

叠氮化物交联剂在水中的光化学

基本信息

批准号：
7623586
负责人：
Christopher M. Hadad
金额：
$ 21.6万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-06-01 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7623586
关键词：
Amino Acids Arts Azides Bacteria Benzophenones Biological Buffers Cell Adhesion Cell surface Chemical Structure Chemicals Chemistry Collaborations Complex Computer Retrieval of Information on Scientific Projects Database Cross-Linking Reagents DNA DNA Polymerase I Databases Environment Escherichia coli Family Foundations Goals Halogens Higher Order Chromatin Structure Ketones Label Laboratories Learning Light Literature Maps Mediating Methodology Methods Microbial Biofilms Nitrogen Nucleic Acids Nucleoproteins Nucleosides Ohio Organic Chemicals Organic Chemistry Organic solvent product Photoaffinity Labels Photochemistry Principal Investigator Property Protein Family Proteins Publications Publishing RNA Reaction Reagent Renaissance Reporting Research Research Personnel Resolution Resources Scientist Series Site Solutions Spectrum Analysis Structural Protein Structure Students Sulfur System Techniques Time Triplet Multiple Birth United States National Institutes of Health Universities Water Work absorption adduct aqueous base chromophore covalent bond crosslink design experience functional group insight macromolecule meetings member nanosecond photolysis professor programs quantum reaction rate research study two-photon

项目摘要

DESCRIPTION (provided by applicant): Azide based reagents are used extensively by NIH supported investigators in photoaffinity labelling and cross-linking studies. Most practitioners utilize a few commercially available azide reagents. It is generally believed that the photochemistry of these reagents in water is similar to that of simple phenyl azides in organic solvents. Preliminary results indicate that this assumption is not generally correct. Thus, we will systematically study the photochemistry of widely used commercially available reagents, simple phenyl azides, cinnamyl, sulfonyl, phoshoryl and acyl azides in water and in buffer solution. We will study these reagents by pico, nano and microsecond time-resolved UV-Vis and IR spectroscopy, which will reveal the lifetimes of the light generated reactive intermediates and absolute reactivities of the intermediates with nucleosides and amino acids. The reactive intermediates will be identified with the aid of CASSCF, CASPT2 and DFT calculations and chemical analysis of reaction mixtures. This work will be done with non-NIH resources. With NIH support a series of new azide precursors will be synthesized, which absorb strongly above 330 nm and reliably produce reactive intermediates in water with lifetimes of 0.1-100 ns. In collaboration with OSU colleague Professor Ming-Daw Tsai we will re-investigate a classic system, the E. coli Klenow Fragment I which has been previously studied with a traditional, commercial reagent (8-azido dATP) and a newly developed reagent to demonstrate that the commercial reagent generates untrustworthy results. In collaboration with a second OSU colleague, Professor Mike Freitas, we will study the labelled Klenow Fragment with high throughput Mass Spectrometric techniques. These new reagents will also be evaluated as cross-linking agents by Professors Pei and Dalbey of The Ohio State University (OSU) to cross-link the PNAG and YidG proteins, respectively. A photo-probe group will be created at OSU bringing together synthetic organic chemists, physical-organic chemists, mass spectrometrists and biochemists.

描述（由申请人提供）：NIH支持的研究人员在光性标记和交联研究中广泛使用了基于叠氮化物的试剂。大多数从业者使用一些市售的叠氮化物试剂。人们普遍认为，这些试剂在水中的光化学类似于有机溶剂中简单苯基的光化学。初步结果表明，这个假设通常不正确。因此，我们将系统地研究广泛使用的市售试剂，简单苯基，肉桂，磺酰基，phoshoryl，phoshoryl和酰基叠氮化物的光化学。我们将通过PICO，NANO和微秒时间分辨的UV-VIS和IR光谱研究来研究这些试剂，这将揭示带有核苷和核苷和氨基酸的中间体的光产生的反应性中间体以及中间体的绝对反应性的寿命。反应性中间体将借助CASSCF，CASPT2和DFT计算以及反应混合物的化学分析。这项工作将使用非NIH资源进行。使用NIH支持一系列新的叠氮化前体，将合成一系列新的叠氮化前体，该前体的吸收高于330 nm，并可靠地在水中产生反应性中间体，其寿命为0.1-100 ns。通过与OSU同事Ming-Daw Tsai合作，我们将重新投资经典的系统，即大肠杆菌Klenow碎片I I，此前曾对传统的商业试剂（8-氮杂DATP）进行了研究，并新开发的试剂证明了商业试剂的商业试剂是不受信任的效果。与第二位OSU同事Mike Freitas合作，我们将研究具有高吞吐量质谱技术的标记的Klenow片段。这些新试剂还将被俄亥俄州立大学（OSU）的PEI和Dalbey教授评估为交联药，分别交联PNAG和YIDG蛋白质。将在OSU上创建一个照片探针组，将合成有机化学家，物理有机化学家，质谱师和生物化学家汇总在一起。