使用乙烯化学物质造异酸键:用于研究异酸酶的活性,对ubiquitin分子进行特定位点的合
Ellen M Valkevich1, Robert G Guenette, Nicholas A Sanchez
1Department of Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin 53706-1322, USA.
Journal of the American Chemical Society
|April 14, 2012
概括
硫-乙烯合 (TEC) 化学可以创建分支的无处不在片. 这种方法使研究人员能够研究ubiquitin的排列如何影响deubiquitinase活性,从而揭示了对蛋白质调节的新见解.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 分子生物学分子生物学
背景情况:
- 蛋白质修饰对于理解生化功能至关重要.
- 乌比基结合在细胞信号通路中起着关键作用.
- 分支的无处不在链参与各种细胞过程.
研究的目的:
- 为了证明乙烯合 (TEC) 化学对蛋白质结合的有用性.
- 为了合成和研究分支ubiquitintrimers的功能.
- 探索乌比奎丁链接导向对二维基基酶 (DUB) 活性的影响.
主要方法:
- 使用TEC化学的特定位点异酸键形成.
- 带有定义链接的分支泛素trimers的合成.
- 使用二维基化酶 (DUB) 分析化链水解的酶定试验.
主要成果:
- 通过TEC化学,成功地在ubiquitin分子之间建立了特定位点的异类键.
- 合成了分支的泛素三元体,使得能够研究链接特异性相互作用.
- 在分支trimers中,ubiquitin单元的定向显著影响了DUB介导的水解,K48C链接裂变被相邻的K6C结合抑制.
结论:
- TEC 是一种强大的工具,用于生成均的无素-蛋白质结合物和链接.
- 分支无处不在链的结构布局决定了DUBs的识别和处理.
- 这些发现表明,分支无素链在链接选择性相互作用和细胞信号传递中起着调节作用.
相关概念视频
01:13The Proteasome
Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3 (ubiquitin...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3 (ubiquitin...
02:18The Proteasome
Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
01:26Protein Modifications in the RER
Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal sequences.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal sequences.
01:24Tagging and Fusion Proteins
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
02:43Peptide Bonds
A peptide bond covalently attaches amino acids through a dehydration reaction. One amino acid's carboxyl group and another amino acid's amino group combine, releasing a water molecule. The resulting bond is the peptide bond. The products that such linkages form are peptides. As more amino acids join this growing chain, the resulting chain is a polypeptide. Each polypeptide has a free amino group at one end. This end has the N-terminal, or the amino-terminal, and the other end has a free...
01:17The Proteasome Structure
The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...
The proteasome is an...