相关实验视频

Updated: Jul 26, 2025

Measuring RAN Peptide Toxicity in C. elegans

Measuring RAN Peptide Toxicity in C. elegans

Published on: April 30, 2020

与C9orf72相关的二重复的毒性是由常用的蛋白质标签修改的

Javier Morón-Oset¹, Lilly Ks Fischer¹, Mireia Carcolé^2,3

¹Max Planck Institute for Biology of Ageing, Cologne, Germany.

Life science alliance

|June 12, 2023

概括

蛋白标签可以改变与C9orf72相关的神经退行性疾病相关的二重复蛋白 (DPR) 的毒性. 研究人员发现,标签可以掩盖或增强DPR毒性,影响临床前研究结果.

科学领域:

神经遗传学神经遗传学
分子毒理学分子毒理学
在Drosophila melanogaster模型中.

背景情况:

在C9orf72基因中的六核酸重复扩张是肌缩侧面硬化症 (ALS) 和前性痴呆症 (FTD) 的主要遗传原因.
这些扩张产生有毒二重复 (DPR) 蛋白质,涉及疾病的发病.
临床前研究经常使用标记的DPR结构,但这些标签对毒性的影响仍然不太清楚.

研究的目的:

系统地研究不同蛋白标签 (mCherry,GFP,FLAG) 对Drosophila模型中C9orf72-关联DPR的毒性的影响.
为了确定蛋白质标签是否影响DPR稳定性,降解和相关的细胞损伤.
评估标签诱导的毒性改变对ALS和FTD临床前研究的影响.

主要方法:

在Drosophila melanogaster.中表达各种标记和未标记的富含氨酸的DPRs (包括GA100).
通过对的表型分析评估DPR毒性.
评估DNA损伤和p62水平作为细胞压力的指标.
对DPR蛋白稳定性和降解途径的分析.

主要成果:

36重复的dprs的mCherry标记增加了毒性,而GA100的mCherry或GFP标记消除了毒性.
FLAG标签降低了GA100的毒性,但不如光标签那么显著.
没有标记的GA100,但没有标记的版本,诱导DNA损伤和p62水平升高,表明细胞效应改变.
光标签影响了GA100蛋白质的稳定性和降解.

结论:

蛋白标签以标签和DPR依赖的方式显著影响DPR毒性.
在使用标记结构的研究中,DPRs,特别是GA的毒性可能被低估.
包括未标记的DPR控制对于准确评估C9orf72相关神经退行性疾病的临床前模型中的毒性至关重要.

更多相关视频

Assays for the Degradation of Misfolded Proteins in Cells

Assays for the Degradation of Misfolded Proteins in Cells

Published on: August 28, 2016

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Published on: July 16, 2021

相关概念视频

Tagging and Fusion Proteins

Tagging 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...

Covalently Linked Protein Regulators

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....