Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506, United States.
Computational Sciences, University of Kentucky, Lexington, Kentucky 40506, United States.
Anal Chem. 2024 Oct 15;96(41):16298-16306. doi: 10.1021/acs.analchem.4c03314. Epub 2024 Oct 2.
We developed a method for receptor encapsulation and single-molecule imaging techniques from neuronal and cardiac tissues, illustrating the method's broad applicability for measuring membrane receptor assembly. Ryanodine receptor 2 (RyR2) is a tetrameric Ca channel governing intracellular Ca dynamics, which is critical for muscle contraction. Employing GFP-RyR2 knock-in mice, we isolated individual receptor proteins in tissue-specific nanovesicles and performed subunit counting analyses to yield quantitative assessment of stoichiometric distributions across different organs. With this method, we explored the potential heterogeneity of brain-derived RyR2, which has been reported to form heteromeric assemblies with other ryanodine receptor isoforms.
我们开发了一种从神经元和心肌组织中包封受体并进行单分子成像技术的方法，该方法说明了其在测量膜受体组装方面的广泛适用性。兰尼碱受体 2（RyR2）是一种四聚体 Ca 通道，可调节细胞内 Ca 动力学，这对于肌肉收缩至关重要。利用 GFP-RyR2 基因敲入小鼠，我们在组织特异性纳米囊泡中分离了单个受体蛋白，并进行亚基计数分析，从而对不同器官的化学计量分布进行定量评估。通过这种方法，我们探讨了脑源性 RyR2 的潜在异质性，据报道，它与其他兰尼碱受体亚型形成异源二聚体。
