在最多三个不同的位置进行精确的蛋白质标记的相互直角无意义抑制系统和结合化学
James S Italia1, Partha Sarathi Addy1, Sarah B Erickson1
1Department of Chemistry , Boston College , 2609 Beacon Street, 246B Merkert Chemistry Center , Chestnut Hill , Massachusetts 02467 , United States.
Journal of the American Chemical Society
|March 27, 2019
概括
科学家们开发了一种新方法,使用直角遗传密码将多达三种不同的非正规氨基酸 (ncAAs) 插入蛋白质中. 这一突破为高级生物应用提供了精确的多部位蛋白质标签.
科学领域:
- 生物化学
- 合成生物学
- 分子生物学
背景情况:
- 在先进的生物应用中,特定位置的蛋白质修饰至关重要.
- 目前将多个非正规氨基酸 (ncAAs) 纳入蛋白质的方法是有限的,最多可以使用两种古老的cAAs.
- 扩大工具包以纳入多个不同的ncAAs对于新型蛋白质工程至关重要.
研究的目的:
- 开发一种系统,将三个不同的非正规氨基酸 (ncAAs) 具体地纳入一个蛋白质中.
- 识别和证明多个生物结合手柄对直角蛋白质进行标记的兼容性.
- 在最多三个不同的位置实现同时对蛋白质进行特定的标记.
主要方法:
- 工程化直角氨基-tRNA合成酶/tRNA对,包括大肠杆菌的三甲基对,与古老的铁和铁甲基对相结合.
- 相互直角的无意义抑制系统被用于整合不同的ncAAs.
- 用于生物结合的化学选择性标记反应包括CRACR,SPAAC和SPIEDAC.
主要成果:
- 通过使用大肠杆菌的三法尼尔对与铁或解对结合,证明了两种不同的ncAAs的成功结合.
- 通过组合三个直角对,同时将三个不同的ncAAs纳入一个单一的蛋白质.
- 展示了CRACR,SPAAC和SPIEDAC的兼容性,用于快速,无催化剂,多部位的蛋白质标签与不同的化学实体.
结论:
- 已经建立了一个用于将多达三种不同的非正规氨基酸 (ncAAs) 纳入蛋白质的新系统.
- 开发的方法允许在多个预定义位置使用兼容的生物结合化学物质进行正交蛋白标记.
- 这种进步显著扩大了合成生物学中的蛋白质工程和功能化能力.
相关概念视频
Conjugated Proteins
27.6K
Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties.
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
27.6K
Conjugated Proteins
3.2K
3.2K
Nonsense-mediated mRNA Decay
11.8K
The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
11.8K
Nonsense-mediated mRNA Decay
3.3K
3.3K
Frost Circles for Different Conjugated Systems
3.6K
The inscribed polygon method is consistent with Hückel’s 4n + 2 rule and helps to learn whether the given cyclic compound is aromatic or not. The compound is stable and aromatic if every bonding molecular orbital (MO) is completely filled with a pair of electrons. However, if the non-bonding or antibonding orbitals are filled with electrons, the compound is unstable and not aromatic. Consider the Frost circle diagrams for cycloalkenes containing 4 to 8 carbons.
3.6K
UV–Vis Spectroscopy of Conjugated Systems
8.4K
Organic compounds with conjugated double bonds show strong absorption features in the UV–visible region of the electromagnetic spectrum attributed to π → π* electronic excitations. Generally, a UV–vis absorption spectrum is recorded as a plot of absorbance vs wavelength. The wavelength of maximum absorbance, which manifests as a peak in the absorption spectrum, is denoted as λmax.
One of the factors influencing λmax is the extent of conjugation in...
One of the factors influencing λmax is the extent of conjugation in...
8.4K


