对感应的STIM1 EF手进行了全面的突变性表征,揭示了对结构和功能至关重要的残留物
Nisha D Kamath1, Kenneth A Matreyek1
1Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.
bioRxiv : the preprint server for biology
|February 3, 2025
概括
我们使用基于细胞的健身测试对706种STIM1 EF手变体进行了功能性表征. 这揭示了STIM1序列变异如何影响信号和疾病,有助于临床解释.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 信号调节基本的细胞过程.
- 储存运行的入 (SOCE) 涉及STIM1感知ER并触发流入.
- STIM1变种可以导致肌肉病变和免疫缺陷等疾病,但很少有特征.
研究的目的:
- 通过评估706个单个氨基酸变体,对STIM1规范EF手域进行功能性表征.
- 了解STIM1序列变化如何影响其在信号传递中的功能.
- 帮助临床解释在患者中发现的STIM1变异.
主要方法:
- 在培养的人类细胞中利用基于STIM1变异过度表达的高通量健身试验.
- 分析了协调残留物和STIM1螺旋体中的突变模式.
- 采用了直角测量方法,包括STIM1寡合化,细胞质流入和细胞应激.
主要成果:
- 在EF手协调残留物中确定了不同的突变模式.
- 在尾行螺旋中发现了一个不变残留的核心,可能抑制STIM1聚合.
- 发现主要的螺旋变体仅在内源性STIM1存在时引起毒性,这表明多元化依赖调节.
结论:
- STIM1规范的EF手序列包含了对SOCE至关重要的复杂角色.
- 细胞毒性测试,结合寡合化和流量测量,增强变体的功能理解.
- 大多数致病性STIM1变体在本试验中没有明显的细胞毒性,这突显了对补充分析的需要.
相关概念视频
Calmodulin-dependent Signaling
5.1K
Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
5.1K
Conserved Binding Sites
4.2K
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
4.2K
Structure of Cadherins
3.3K
The cadherins were one of the first cell adhesion molecules discovered; the term “cadherins” is based on their calcium-dependent adhering properties. The first cadherins discovered on the epithelial, neuronal, and placental cells were named E-cadherin, P-cadherin, and N-cadherin, respectively. These classical cadherins share sequence and structural similarities. Other cadherins, including those involved in cell signaling, are grouped into non-classical cadherins. This...
3.3K
Overview of Secretory Vesicles
8.3K
Secretory vesicles, also known as dense core vesicles (DCVs), are membrane-bound vesicles that transport secretory proteins, such as hormones or neurotransmitters. Regulated secretory vesicles transport proteins from the trans-Golgi network to the exterior of the cell. Proteins present in regulated secretory vesicles are required to be rapidly exocytosed in large amounts upon a specific stimulus.
Various proteins regulate the aggregation of molecules inside the secretory vesicles. Chromogranins...
Various proteins regulate the aggregation of molecules inside the secretory vesicles. Chromogranins...
8.3K


