通过蛋白质组方法对胰岛素及其从S9肝脏中的代谢物中的二硫化键进行全面的表征
Chenxi Wang1, Xinyuan Ye1, Siying Zheng1
1Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030, China.
Journal of proteome research
|October 14, 2025
概括
这项研究引入了一种新的蛋白质组策略,用于准确地绘制治疗性和蛋白质 (TPP) 和其代谢物的二硫化键,克服分析过程中混合键的挑战.
科学领域:
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
- 分析化学 分析化学
背景情况:
- 二硫化键对于治疗性和蛋白质 (TPP) 的结构和功能至关重要.
- 在TPP及其代谢物中准确地绘制二硫化物键图是具有挑战性的,原因是目前软件的键盘编码和局限性.
研究的目的:
- 开发一个全面的蛋白质组学策略,以表征TPP及其代谢物中的二硫化物键.
- 解决现有软件在识别具有非原生二硫化键的代谢物的局限性.
主要方法:
- 使用pLink软件开发了一个蛋白质组策略.
- 样品的制备过程包括在0°C的胃消化,然后是液体染色学高分辨率质谱学 (LC-HRMS).
- 代谢物验证涉及使用甲基氧酸 (mCPBA) 的减少,化和氧化.
主要成果:
- 胰岛素中的所有二硫化键都被识别出来,没有混.
- 从鼠肝S9中鉴定出118种胰岛素代谢物,其中41种呈现出二硫化键重组.
- 该策略成功验证了新陈代谢的胰岛素与重新混合的二硫化键.
结论:
- 开发的战略使TPP及其代谢物的可靠和全面的表征成为可能,包括那些具有重组的二硫化物键的代谢物.
- 这种方法对于TPPs的临床前和临床开发至关重要.
更多相关视频
09:37Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture
Published on: May 2, 2019
10.7K
07:16Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
2.1K
相关概念视频
Protein and Protein Structure
86.8K
Proteins are one of the most abundant organic molecules in living systems and have the most diverse range of functions of all macromolecules. Proteins may be structural, regulatory, contractile, or protective. They may serve in transport, storage, or membranes; or they may be toxins or enzymes. Their structures, like their functions, vary greatly. They are all, however, amino acid polymers arranged in a linear sequence.
A protein's shape is critical to its function. For example, an enzyme...
A protein's shape is critical to its function. For example, an enzyme...
86.8K
Protein Modifications in the RER
6.9K
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...
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...
6.9K
Insulin: Biosynthesis, Chemistry, and Preparation
1.2K
The endoplasmic reticulum (ER) of pancreatic β-cells synthesizes preproinsulin, which consists of a signal peptide, A and B chains, and a C-peptide. Preproinsulin is then cleaved and folded into proinsulin, which translocates to the Golgi apparatus for sorting and packaging into secretory granules. In these granules, enzymatic clipping generates insulin and C-peptide.
Damage or functional impairment of β-cells inhibits insulin production, leading to diabetes. Diabetes treatment...
Damage or functional impairment of β-cells inhibits insulin production, leading to diabetes. Diabetes treatment...
1.2K
