带有C端标签的细菌J域与DnaK的基质结合域联系,并隔离护送活动
Brock Nelson1, Nathan Soper1, Tania J Lupoli1
1Department of Chemistry, New York University, New York, 10003, USA.
Chembiochem : a European journal of chemical biology
|August 9, 2023
概括
将特定标签添加到菌根菌的J蛋白中,可以增强它们与DnaK伴侣的相互作用. 这种修饰影响了细胞蛋白质稳定,并为研究伴侣-辅助因子动态提供了新的途径.
科学领域:
- 分子生物学分子生物学
- 微生物学 微生物学
- 蛋白质相互作用 蛋白质相互作用
背景情况:
- 像DnaK这样的分子伴侣和像J蛋白这样的辅因子对于维持细胞蛋白质平衡 (蛋白质平衡) 至关重要.
- 在Mycobacterium结核病中,DnaK-DnaJ相互作用对细菌的生存至关重要,并代表潜在的治疗点.
- J-蛋白的J域与DnaK相互作用,但其他域的作用不太清楚.
研究的目的:
- 调查切断的真菌DnaJ1和DnaJ2J域上的C端标签如何影响它们与DnaK的相互作用.
- 探索这些修饰的J域的潜力,作为研究伴侣-辅助因子关系的工具.
主要方法:
- 菌根DnaJ1和DnaJ2的截断的N终端J域与C终端标签 (例如His6) 进行了工程设计.
- 分析了这些修改后的J域与DnaK的结合,重点关注DnaK的基质结合域内的相互作用.
- 评估了在蛋白质毒性压力下的真菌细菌细胞中表达这些修饰的J域的功能影响,特别是与DnaK抑制剂结合使用.
主要成果:
- 他的6个标签独特地促进了J域与DnaK的C端基质结合域的额外结合.
- 其他测试的C端标签,包括已知的DnaK相互作用,没有诱导类似的增强结合.
- 在蛋白质毒性压力下,mycobacteria中C终端修饰的DnaJJ域的表达损害了伴侣活动,这种效应被DnaK抑制剂放大.
结论:
- C终端的His6标签可以在J域和DnaK之间创建新的交互站点.
- 基因可编码的,改造的J蛋白变体可以开发来调节伴侣活动.
- 这些发现为剖析各种生物系统中伴侣-辅助因子相互作用提供了新的工具.
更多相关视频
07:14Author Spotlight: Exploring Heat Shock Proteins in Malaria and Tuberculosis Infections
Published on: March 8, 2024
1.4K
10:24Defining Hsp33's Redox-regulated Chaperone Activity and Mapping Conformational Changes on Hsp33 Using Hydrogen-deuterium Exchange Mass Spectrometry
Published on: June 7, 2018
8.7K
相关概念视频
Bacterial Protein Maturation
36
Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
36
Single-Strand DNA Binding Proteins
14.2K
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...
14.2K
Molecular Chaperones and Protein Folding
18.0K
The native conformation of a protein is formed by interactions between the side chains of its constituent amino acids. When the amino acids cannot form these interactions, the protein cannot fold by itself and needs chaperones. Notably, chaperones do not relay any additional information required for the folding of polypeptides; the native conformation of a protein is determined solely by its amino acid sequence. Chaperones catalyze protein folding without being a part of the folded protein.
The...
The...
18.0K
Conservation of Protein Domains Over Different Proteins
10.9K
Protein domains are small structurally independent units that are part of a single amino acid chain. Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
10.9K
Assembly of Signaling Complexes
5.8K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
5.8K
Cooperative Binding of Transcription Regulators
6.5K
Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome. Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
6.5K
