通过红外光谱学解决的DNA寡核酸脱杂的依赖序列机制
Paul J Sanstead1, Paul Stevenson1,2, Andrei Tokmakoff1
1Department of Chemistry, Institute for Biophysical Dynamics, and James Franck Institute, The University of Chicago , 929 East 57th Street, Chicago, Illinois 60637, United States.
Journal of the American Chemical Society
|August 14, 2016
概括
DNA寡核酸杂化与简单的两态解离有所不同. 红外光谱显示基对特定的机制, 显示终端脱光, 而不是循环,
科学领域:
- 分子生物学
- 生物物理
- 光谱学
背景情况:
- 在生物学和纳米技术中,氧核化非常重要.
- 没有完全理解DNA杂交的分子机制和动态.
- 现有的模型通常假定一个简单的两种状态分离路径.
研究的目的:
- 开发和应用一种新的策略来表征DNA寡核酸的基对特异性热解离机制.
- 调查从假设的两个状态解离模型的偏差.
- 阐明序列和基对组成在杂交动态中的作用.
主要方法:
- 使用稳定状态和时间分辨率的红外光谱 (FTIR和2D-IR).
- 使用格子模型来对实验数据进行结构特定的解释.
- 研究了自补的10个基对DNA序列与不同的GC基对放置.
主要成果:
- 在DNA复合体中观察到与简单的两态解离行为的显著偏差.
- 证明双重解离主要是通过终端基层脱而不是循环形成.
- 量化解离时间尺度:在磨损时70-100 ns,在融化温度附近完全解离时10-30 μs.
结论:
- 这项研究提供了基对特异性的DNA寡核酸解离机制的理解.
- 终端脱被认为是主要的途径,挑战了简化的两种状态模型.
- 核基序列在双重解离过程中极大地影响了脱状态的性质.
相关概念视频
IR Spectrum Peak Broadening: Hydrogen Bonding
2.1K
The vibrational frequency of a bond is directly proportional to its bond strength. As a result, stronger bonds vibrate at higher frequencies, while weaker bonds vibrate at lower frequencies. The stretching vibration of the strong O–H bond in alcohols and phenols (very dilute solution or gas phase) appears as a sharp peak at 3600–3650 cm−1.
However, the extent of hydrogen bonding influences the observed stretching frequency and band broadening. Intermolecular or intramolecular...
However, the extent of hydrogen bonding influences the observed stretching frequency and band broadening. Intermolecular or intramolecular...
2.1K
Maxam-Gilbert Sequencing
13.5K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
13.5K
Homologous Recombination
65.0K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
65.0K
IR and UV–Vis Spectroscopy of Aldehydes and Ketones
7.8K
Infrared spectroscopy, also known as vibrational spectroscopy, is mainly used to determine the types of bonds and functional groups in molecules. In aldehydes and ketones, the carbonyl (C=O) bond shows an absorption around 1710 cm-1. The C=O bond vibration of an aldehyde occurs at lower frequencies than that of a ketone. In addition to the C=O absorption in an aldehyde, the aldehydic C–H bond also gives two peaks in the 2700–2800 cm-1 range. This absorption, coupled with the...
7.8K
¹³C NMR: ¹H–¹³C Decoupling
2.0K
The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
2.0K
Single-Strand DNA Binding Proteins
17.1K
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
17.1K


