Department of Cell Biology and Neuroscience, Graduate School of Biomedical Sciences, School of Osteopathic Medicine, Rowan University, Stratford, NJ 08084, USA.
Department of Rehabilitation Medicine, NeuroMusculoskeletal Institute, School of Osteopathic Medicine, Rowan University, Stratford, NJ 08084, USA.
Molecules. 2021 Jan 5;26(1):227. doi: 10.3390/molecules26010227.
S100B is a calcium-binding protein that governs calcium-mediated responses in a variety of cells-especially neuronal and glial cells. It is also extensively investigated as a potential biomarker for several disease conditions, especially neurodegenerative ones. In order to establish S100B as a viable pharmaceutical target, it is critical to understand its mechanistic role in signaling pathways and its interacting partners. In this report, we provide evidence to support a calcium-regulated interaction between S100B and the neuronal calcium sensor protein, neurocalcin delta both in vitro and in living cells. Membrane overlay assays were used to test the interaction between purified proteins in vitro and bimolecular fluorescence complementation assays, for interactions in living cells. Added calcium is essential for interaction in vitro; however, in living cells, calcium elevation causes translocation of the NCALD-S100B complex to the membrane-rich, perinuclear trans-Golgi network in COS7 cells, suggesting that the response is independent of specialized structures/molecules found in neuronal/glial cells. Similar results are also observed with hippocalcin, a closely related paralog; however, the interaction appears less robust in vitro. The N-terminal region of NCALD and HPCA appear to be critical for interaction with S100B based on in vitro experiments. The possible physiological significance of this interaction is discussed.
S100B 是一种钙结合蛋白,可调节多种细胞(尤其是神经元和神经胶质细胞)中的钙介导反应。它也被广泛研究作为几种疾病状况,特别是神经退行性疾病的潜在生物标志物。为了将 S100B 确立为可行的药物靶点,了解其在信号通路中的机制作用及其相互作用伙伴至关重要。在本报告中,我们提供了证据支持 S100B 与神经元钙传感器蛋白神经钙蛋白 δ(neurocalcin delta)之间在体外和活细胞中的钙调节相互作用。膜覆盖测定法用于测试体外纯化蛋白之间的相互作用,而双分子荧光互补测定法用于活细胞中的相互作用。体外相互作用需要额外的钙;然而,在活细胞中,钙的升高导致 NCALD-S100B 复合物易位到富含膜的、核周反式高尔基体网络,这表明该反应独立于神经元/神经胶质细胞中发现的特殊结构/分子。与密切相关的同源物 hippocalcin 也观察到类似的结果;然而,体外相互作用似乎不太强。基于体外实验,NCALD 和 HPCA 的 N 端区域似乎对与 S100B 的相互作用至关重要。讨论了这种相互作用的可能生理意义。