使用分离的酵母微粒体进行体外同型内质网（ER）膜融合测定
In Vitro Homotypic ER Membrane Fusion Assay Using Isolated Yeast Microsomes.
作者信息
Jang Eunhong, Jun Youngsoo
机构信息
School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, South Korea.
出版信息
Methods Mol Biol. 2025;2887:167-173. doi: 10.1007/978-1-0716-4314-3_11.
Cell-free in vitro assays offer several advantages for elucidating molecular mechanisms underlying various biological processes. Here, we describe a simple and quantitative in vitro assay using isolated yeast microsomes to measure homotypic ER membrane fusion. In this assay, membrane fusion between ER microsomes is monitored by reconstitution of luciferase activity from split luciferase fragments. Our findings reveal that homotypic ER membrane fusion requires not only Sey1p, the yeast atlstin, but also ER-resident SNAREs, such as Sec22p and Sec20p, in Saccharomyces cerevisiae.
无细胞体外分析对于阐明各种生物学过程背后的分子机制具有诸多优势。在此，我们描述了一种简单且定量的体外分析方法，该方法使用分离的酵母微粒体来测量同型内质网（ER）膜融合。在这种分析中，通过从分裂的荧光素酶片段重构荧光素酶活性来监测内质网微粒体之间的膜融合。我们的研究结果表明，在酿酒酵母中，同型内质网膜融合不仅需要酵母atlastin Sey1p，还需要内质网驻留的SNARE蛋白，如Sec22p和Sec20p。
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