Jove
Visualize
联系我们
JoVE
x logofacebook logolinkedin logoyoutube logo
关于 JoVE
概览领导团队博客JoVE 帮助中心
作者
出版流程编辑委员会范围与政策同行评审常见问题投稿
图书馆员
用户评价订阅访问资源图书馆顾问委员会常见问题
研究
JoVE JournalMethods CollectionsJoVE Encyclopedia of Experiments存档
教育
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab Manual教师资源中心教师网站
使用条款与条件
隐私政策
政策

相关概念视频

Protein-protein Interfaces02:04

Protein-protein Interfaces

Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
Protein Networks02:26

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,...
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...

您也可能阅读

相关文章

通过共同作者、期刊和引用图与本文相关的文章。

排序
Same author

A first-in-pediatric study of ALRN-6924, a novel stapled-peptide dual MDM2/MDMX inhibitor, for children with advanced hematologic and solid malignancies.

Clinical cancer research : an official journal of the American Association for Cancer Research·2026
Same author

Transient p53/p21 activation selectively protects healthy human hair follicles and their stem cells from chemotherapy.

The Journal of clinical investigation·2026
Same author

Discovery of Sulanemadlin (ALRN-6924), the First Cell-Permeating, Stabilized α-Helical Peptide in Clinical Development.

Journal of medicinal chemistry·2023
Same author

Exploring new targets and chemical space with affinity selection-mass spectrometry.

Nature reviews. Chemistry·2023
Same author

Corrigendum to "Impact of enzyme concentration and residence time on apparent activity recovery in jump dilution analysis" [Anal. Biochem. 416 (2011) 206-210].

Analytical biochemistry·2022
Same author

Correction: Phase I Trial of ALRN-6924, a Dual Inhibitor of MDMX and MDM2, in Patients with Solid Tumors and Lymphomas Bearing Wild-type TP53.

Clinical cancer research : an official journal of the American Association for Cancer Research·2022

相关实验视频

Updated: Jul 6, 2026

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
12:43

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study

Published on: July 27, 2016

在化合物混合物中,一种一般技术来排列蛋白质 - 配体结合的亲缘关系,并确定异质与直接结合位点的竞争.

D Allen Annis1, Naim Nazef, Cheng-Chi Chuang

  • 1NeoGenesis Pharmaceuticals Inc., 840 Memorial Drive, Cambridge, MA 02139, USA. aannis@neogenesis.com

Journal of the American Chemical Society
|November 26, 2004
PubMed
概括

我们开发了一种亲和性选择质谱法 (AS-MS) 方法来排列蛋白质-连接体结合亲和性,并根据结合位点对连接体进行分类. 这个工具通过有效地识别和优化用于疾病标的化合物,有助于药物发现.

更多相关视频

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

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

相关实验视频

Last Updated: Jul 6, 2026

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
12:43

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study

Published on: July 27, 2016

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

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

科学领域:

  • 生物化学 生物化学
  • 化学生物学 化学生物学
  • 药物发现 药物发现 药物发现

背景情况:

  • 组合化学为药物发现提供了巨大的潜力,但需要有效的工具来识别和优化化合物.
  • 目前用于蛋白质 - 配体相互作用分析的方法可能是资源密集的,缺乏全面的结合位点分类.
  • 开发通用,高效的工具对于利用药物开发中的以多样性为导向的合成至关重要.

研究的目的:

  • 引入和验证一种新的亲和力选择-质谱法 (AS-MS) 方法,用于蛋白质-配体亲和力排名和结合位点分类.
  • 在药物发现中,为各种蛋白标和配体类提供适用于各种蛋白标的多功能工具.
  • 为了能够在不需要复杂的测试的情况下,对结合性亲缘关系和合作性的定量估计.

主要方法:

  • AS-MS方法涉及从配体池中选择蛋白质结合化合物的亲和力,使用竞争对手的配体.
  • 通过染色学将未结合的连接物从蛋白质-连接物复合物分离出来.
  • 通过质谱法 (MS) 分离和识别/定量联体,以确定亲和力和结合部位.

主要成果:

  • 证明AS-MS对各种蛋白质类的成功应用,包括激酶 (Akt-1,Zap-70) 和GPCR (M(2) 肌肉性乙胆受体).
  • 确定了新的ATP竞争性和全性抑制剂,以及以前未公开的抗剂.
  • 验证了该方法在量化估计结合性亲缘关系和结合性合作性方面的能力.

结论:

  • AS-MS方法是一种强大的,无标签的技术,用于蛋白质-配体亲和度排名和结合位点分类.
  • 它不需要任何化合物或蛋白质修饰,可以在溶液中进行.
  • 该方法非常适用于高通量查,优化和药物发现中的结构-活性关系研究.