膜拓和高估蛋白质集群在单分子定位显微镜 - 识别和纠正
Jeremy Adler1, Kristoffer Bernhem2, Ingela Parmryd3
1Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Communications biology
|July 1, 2024
概括
等离子体膜蛋白质集群可能是膜拓学的工件. 研究人员开发了一种使用膜标记物的方法,以区分真实蛋白质集群和虚假的蛋白质集群,揭示了地形.
科学领域:
- 细胞生物学 细胞生物学
- 生物物理学的生物物理.
- 显微镜的使用方法
背景情况:
- 单分子局部化显微镜 (SMLM) 表明大多数血膜蛋白质形成集群.
- 解释SMLM数据需要区分真正的蛋白质聚合与人工物.
- 膜地形的变化可能会产生假阳性聚类信号.
研究的目的:
- 调查膜地形的变化是否会导致膜蛋白的明显聚类.
- 开发和验证一种方法来区分真正的蛋白质集群和地形诱导的文物.
- 重新评估特定膜蛋白,转移素受体 (TfR) 和CD59的聚类,考虑膜拓.
主要方法:
- 使用了双色活细胞SMLM.
- 膜探头DiI与TfR和CD59.9一起使用.
- 分析包括对相关性,对相关性-光激活局部化显微镜和最近邻近分析.
- 图像分析结合了基于DiI的膜拓校正.
主要成果:
- 标准分析表明TfR,CD59和DiI的聚类.
- 在使用DiI信号对膜地形进行校正后,CD59集群不再被检测到,这表明它们是文物.
- 即使经过地形校正,TfR集群仍然可以检测到,这表明真正的集群.
结论:
- 膜地形变化可以导致标准集群分析方法的假阳性结果.
- 使用膜标记器的地形校正方法有效地区分真假蛋白质集群.
- 虽然CD59集群似乎是膜拓学的工件,但TfR表现出真正的集群.
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