Dynamics of myristoylated amino-terminal domain of Src in the plasma membrane of mechano-stimulated cells studied by single molecule imaging

单分子成像研究机械刺激细胞质膜中 Src 肉豆蔻酰化氨基末端结构域的动态

• 批准号: 23612002
• 负责人: KOBAYASHI Takeshi
• 金额: $ 3.49万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23612002/
• 关键词: シグナル伝達; 生物物理; １分子計測; 1分子計測

Src is a critical signaling molecule for mechanotransduction. Mechanical stimuli applied on the cell can induce a rapid Src activation, but it remains largely unknown how mechanical stimuli activates Src molecules. Here, we observed fluorescently labeled myristoylated amino terminus of Src in the plasma membrane of living cells at the single molecule level to examine the role of lipid moiety of the amino terminus of Src on its dynamics and activation. Most probes were observed to diffuse rapidly in the membrane, but that significant fraction of the molecules exhibited temporal immobilization. We also found that the immobilization of them occurred frequently in the proximity of focal adhesions (FAs), and that it was mechano- and cholesterol-dependent. These results suggest that lipid moiety of the amino terminus of Src may play a critical role in the dynamics and activation of Src molecules around FAs through the interaction with lipid micro domains of the plasma membrane.

SRC是机械转导的关键信号分子。施加在细胞上的机械刺激可以诱导Src的快速激活，但机械刺激如何激活Src分子在很大程度上仍不清楚。在这里，我们在单分子水平上观察到活细胞质膜中的Src的肉豆蔻酰化氨基末端的荧光标记，以检测其氨基末端的脂质部分对其动力学和激活的作用。观察到大多数探针在膜中迅速扩散，但有相当一部分分子表现出暂时的固定。我们还发现，它们的固定经常发生在局灶性粘连(FAs)附近，而且它是机械和胆固醇依赖的。这些结果表明，Src氨基末端的脂质部分可能通过与质膜脂质微区的相互作用，在FA周围的Src分子的动态和激活中起关键作用。

项目成果

Sensing the substrate rigidity: possible roles of stress fibers, focal adhesion and mechanosensitive channel

传感基底刚性：应力纤维、粘着斑和机械敏感通道的可能作用

• 发表时间: 2011
• 作者: Kobayashi T;Tanaka M;Takeda Y;Ito G;Sokabe M
• 通讯作者: Sokabe M

名古屋大学大学院医学系研究科・医学部医学科_研究室紹介_細胞科学_細胞生物物理学

名古屋大学大学院医学研究科/医学部医学部_实验室介绍_细胞科学_细胞生物物理学

Cellular Substrate Rigidity Sensing: Possible Role of Mechanosensitive Ion Channels with Stress Fibers and Focal Adhesions

细胞基质刚性传感：机械敏感离子通道与应力纤维和焦点粘附的可能作用

• 发表时间: 2012
• 作者: Kobayashi T;Sokabe M
• 通讯作者: Sokabe M

Cooperation of p40phox with p47phox for Nox2 activation during FcR-mediated phagocytosis --- Mechanism for acquisition of p40phox PI(3)P binding---

p40phox 与 p47phox 合作在 Fc 期间激活 Nox2

• DOI: 10.1074/jbc.m111.237289
• 发表时间: 2011
• 期刊: J. Biol. Chem.
• 作者: Ueyama T;Nakakita J;Nakamura T;Kobayashi T;Kobayashi T;Son J;Sakuma M;Sakaguchi H;Leto TL;Saito N
• 通讯作者: Saito N

Sensing the substrate rigidity : possible roles of stress fibers, focal adhesion and mechanosensitive channel

传感基底刚性：应力纤维、粘着斑和机械敏感通道的可能作用

• 发表时间: 2011
• 作者: Kobayashi T;Tanaka M;Takeda Y;Ito G;Sokabe M
• 通讯作者: Sokabe M