相关实验视频

Updated: Jun 12, 2025

Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens

Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens

Published on: February 6, 2017

使用交叉连接质谱法确定T细胞受体结合方向和-HLA相互作用

Thomas Powell¹, Vijaykumar Karuppiah¹, Saher Afshan Shaikh¹

¹Immunocore Limited, Abingdon, United Kingdom.

The Journal of biological chemistry

|March 28, 2025

概括

确定T细胞受体 (TCR) 与-HLAs的结合方向至关重要. 一种新的交联联质谱法 (XL-MS/MS) 方法有效地解读了TCR-pHLA相互作用和结合几何.

科学领域:

免疫学免疫学免疫学
结构生物学结构生物学
生物化学生物化学

背景情况:

T细胞受体 (TCRs) 对于适应性免疫至关重要,识别-人白细胞抗原 (pHLA) 复合体.
适当的TCR-pHLA结合方向对于T细胞激活至关重要;错误的方向导致非功能性T细胞.
确定TCR-pHLA结合几何学的传统方法,例如晶体结构的确定,可能会耗费大量时间.

研究的目的:

引入和验证一个交叉连接联质谱法 (XL-MS/MS) 方法,以阐明TCR-pHLA结合方向.
为TCR-pHLA相互作用提供一个更快,更简单的替代现有的结构确定技术.

主要方法:

应用交叉连接联质谱法 (XL-MS/MS) 方法来研究TCR-pHLA复合体.
在TCR-pHLA接口内定位交叉链接点到特定的氨基酸残留物.
通过结构预测和精选TCR-pHLA复合物的晶体结构确定,验证XL-MS/MS发现.

主要成果:

通过XL-MS/MS方法,成功地区分了规范和反向TCR与pHLA的结合方向.
交叉链接网站提供了有关TCR-pHLA接口的分子相互作用的详细信息.
结构预测和实验晶体结构证实了由XL-MS/MS确定的结合方向.

结论:

开发的XL-MS/MS方法提供了一种高效且易于使用的方法来确定TCR-pHLA结合方向.
这种技术有助于更深入地了解T细胞受体相互作用,这对免疫反应和治疗发展至关重要.

关键词:

具有约束力的方向导向.交叉链接是指交叉链接.标本地图绘制标本地图绘制质谱测量质谱测量质谱测量质谱测量质量测量质谱测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量反向绑定的反向绑定硫酸-SDA 的使用.

更多相关视频

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay

Published on: October 30, 2015

相关概念视频

Peptide Identification Using Tandem Mass Spectrometry

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...

MALDI-TOF Mass Spectrometry

MALDI-TOF Mass Spectrometry

Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.
Matrix-assisted laser desorption ionization (MALDI) is a commonly...