综合方法确定Robo1的肝硫酸配体需求
Chengli Zong, Rongrong Huang, Eduard Condac
1Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, and Bijvoet Center for Biomolecular Research, Utrecht University , Universiteitsweg 99, 3584 CG Utrecht, The Netherlands.
Journal of the American Chemical Society
|September 10, 2016
概括
研究人员开发了一种新的方法来确定人体圆周受体1 (Robo1) 等蛋白质的肝酸硫酸 (HS) 结合要求. 这种方法揭示了对HS连接体相互作用至关重要的特定结构特征,影响细胞迁移.
科学领域:
- 生物化学
- 葡萄糖生物学
- 分子生物学
背景情况:
- 肝酸硫酸盐 (HS) 结合蛋白在各种生物过程中起着至关重要的作用,包括发育和细胞信号传递.
- 了解HS配体的特定结构要求对于解读蛋白质-碳水化合物相互作用至关重要.
- 人类圆周受体1 (Robo1) 是一个关键的发育蛋白质,其HS结合特性尚未完全表征.
研究的目的:
- 建立一个综合的方法来确定HS结合蛋白的HS配体要求.
- 为了阐明人类Robo1识别的HS的特定结构特征.
- 研究Robo1-HS相互作用对内皮细胞迁移的功能后果.
主要方法:
- 自然HS的部分酶降解,然后进行尺寸排除净化和亲和度丰富.
- 水性相互作用色谱-高分辨率质谱 (HILIC-HRMS) 用于结构性确定.
- 用于结构-活性关系 (SAR) 研究的定义HS寡糖的模块化化学合成.
- 表面等离子共振 (SPR) 用于结合亲和度测量.
主要成果:
- 机器人1表现出一种特定的HS八糖结构子集.
- 详细的结构分析确定了对Robo1结合至关重要的硫化模式.
- 在C-6位置的硫酸对于高亲和度结合至关重要,而C-2硫化则减少结合.
- 为Robo1成功制备和验证了一种高亲和度的合成八糖联体.
- 由Slit2-Robo1信号引起的内皮细胞迁移减少.
结论:
- 开发的综合方法对于表征HS连接物相互作用是有效的.
- Robo1的HS结合具有高度特异性,定义的硫化模式决定了亲和力.
- 了解这些相互作用可以了解Robo1在发育过程和细胞迁移中的作用.
- 这些发现对设计针对Robo1介导途径的基于HS的疗法有影响.
相关概念视频
Ligand Binding Sites
15.6K
Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
15.6K
Ligand Binding Sites
9.0K
9.0K
Assembly of Signaling Complexes
7.0K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
7.0K
Glycosaminoglycans
7.5K
Glycosaminoglycans (GAGs), also known as mucopolysaccharides, are long and linear polymers comprising of specific repeating disaccharides - the amino sugar that can be N-acetylglucosamine or N-acetylgalactosamine, and a uronic acid that is usually glucuronic acid or iduronic acid.
GAGS are found in the extracellular matrix of vertebrates, invertebrates, and bacteria. Due to their polar nature they attract water, and serve as excellent lubricants or shock absorbers in an animal body.
Hyaluronic...
GAGS are found in the extracellular matrix of vertebrates, invertebrates, and bacteria. Due to their polar nature they attract water, and serve as excellent lubricants or shock absorbers in an animal body.
Hyaluronic...
7.5K
Ligand Binding and Linkage
5.9K
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
5.9K
Ligand Binding and Linkage
4.2K
4.2K


