Dusica Maysinger, Issan Zhang, Hao Yuan, Vlasta Bonačić-Koutecký, Željka Sanader Maršić and Rodolphe Antoine
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引用次数: 0
摘要
本研究的目的是确定是否可以在表达胰岛素受体的活细胞中观察到金-金-金(III)偶联胰岛素,以及胰岛素信号传导的关键步骤是否不受干扰。Au(III)与胰岛素的三个酪氨酸配位,并通过与这些酪氨酸配位形成金属离子配位胰岛素模板而具有很强的稳定性。我们结合理论和实验方法,包括计算分析、免疫细胞化学和功能分析来分析这种结构的细胞效应。本研究结果表明,胰岛素- au (III)偶联物不会损害胰岛素信号传导和溶酶体和脂滴下游代谢功能的几个步骤。也就是说,信号激酶蛋白激酶B (Akt1)和丝裂原活化蛋白激酶(ERK1/2)的磷酸化程度与未修饰的胰岛素相当。免疫细胞化学也显示核转录因子EB (TFEB)的减少,这是溶酶体生物发生的重要调节因子。荧光共标记溶酶体LysoTracker和脂滴BODIPY 493/503显示细胞器形态正常,脂滴含量略有增加。胰岛素-金(III)的发光特性可以将生化和成像研究结合起来,用于小胶质细胞以外的人类细胞。
Luminescent insulin–Au(iii) conjugate retains insulin biological properties in human microglia
The aims of this study were to establish if insulin conjugated with auric gold Au(III) can be visualized in living cells expressing insulin receptors, and if key steps in insulin signaling are undisturbed. Au(III) is coordinated with three tyrosines of insulin and is strongly stabilized by coordination with these tyrosines into a metal-ion coordinated insulin template. We combined theoretical and experimental approaches, including computational analyses, immunocytochemistry and functional assays to analyze the cellular effects of this construct. Results in this study show that the insulin–Au(III) conjugate does not impair several steps in insulin signaling and downstream metabolic functions in lysosomes and lipid droplets. Namely, the signaling kinases protein kinase B (Akt1) and mitogen-activated protein kinases (ERK1/2) are phosphorylated to a comparable extent as with unmodified insulin. Immunocytochemistry also shows a comparable decrease in nuclear transcription factor EB (TFEB), an essential regulator of lysosomal biogenesis. Fluorescence co-labelling of lysosomes with LysoTracker and lipid droplets with BODIPY 493/503 shows normal organellar morphology and slightly enhanced lipid droplet content. The luminescent properties of insulin–Au(III) could serve to combine biochemical and imaging studies in human cells beyond microglia.
期刊介绍:
Nanoscale is a high-impact international journal, publishing high-quality research across nanoscience and nanotechnology. Nanoscale publishes a full mix of research articles on experimental and theoretical work, including reviews, communications, and full papers.Highly interdisciplinary, this journal appeals to scientists, researchers and professionals interested in nanoscience and nanotechnology, quantum materials and quantum technology, including the areas of physics, chemistry, biology, medicine, materials, energy/environment, information technology, detection science, healthcare and drug discovery, and electronics.