相关实验视频

Updated: Jun 23, 2025

Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology

Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology

Published on: July 12, 2014

通过HaloTag丰富质谱法绘制脂肪细胞互动原子网络的映射

Junshi Yazaki^1,2, Takashi Yamanashi^1,3,4, Shino Nemoto⁵

¹Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, 230-0045, Japan.

Biology methods & protocols

|June 17, 2024

概括

本研究介绍了基于HaloTag的亲和力净化质谱 (HaloMS) 用于高通量蛋白相互作用的发现. HaloMS 能够快速捕获新表达的蛋白质,从而促进了对细胞网络和蛋白质固体测量的理解.

科学领域:

蛋白质组学是指蛋白质组学
系统生物学系统生物学
分子生物学分子生物学

背景情况:

在体内绘制蛋白相互作用复合体的地图对于理解细胞网络至关重要.
在蛋白质组学中,确定蛋白质相互作用固体测量仍然是一个重要的技术挑战.
人工智能 (AI) 为复杂的生物数据分析提供了潜在的解决方案.

研究的目的:

开发一种高通量方法,用于发现蛋白质相互作用及其固态度.
应用开发的方法来研究人类脂肪细胞中的蛋白质复合体.
在脂肪细胞分化过程中揭示新型转录因子 (TF) -蛋白相互作用.

主要方法:

基于HaloTag的亲和力净化质谱 (HaloMS) 的开发,用于高通量蛋白相互作用的发现.
应用HaloMS来捕获和分析人类脂肪细胞中17种调节蛋白质的蛋白质复合体.
使用体外拉下测试和基于AI的预测工具验证脂肪细胞互动组网络.

主要成果:

HaloMS成功实现了新表达蛋白质的快速捕获和分析,克服了传统分析的局限性.
脂肪细胞互动组网络被绘制和验证,为蛋白质复杂相互作用提供了洞察力.
识别之前未知的转录因子 (TF) -参与脂肪细胞分化中的蛋白质复合体.

结论:

HaloMS是一个强大的,高吞吐量工具,用于蛋白质相互作用的发现和体内静脉测量分析.
该研究揭示了新的TF蛋白复合体,有助于更深入地了解人类脂肪细胞网络和途径集成.
这种方法推进了蛋白质组学和系统生物学领域,通过提供一个强大的方法来绘制复杂的生物相互作用.

关键词:

在 HaloMS 中使用 HaloMS.脂肪细胞的脂肪细胞.交互映射交互映射交互映射网络分析网络分析

更多相关视频

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Published on: November 17, 2019

Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry

Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry

Published on: May 17, 2016

相关概念视频

Protein Networks

Protein Networks

An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...

Proteomics

Proteomics

A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...

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...