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揭示了酵母膜蛋白Pma1的扩散状态,以及标记方法如何改变扩散行为的方法

Mary Lou P Bailey^1,2, Susan E Pratt^1,3, Michael Hinrichsen⁴

¹Integrated Graduate Program in Physical and Engineering Biology, Yale University, New Haven, CT, 06511, USA.

The European physical journal. E, Soft matter

|June 9, 2023

概括

标签方法对酵母膜蛋白Pma1有很大影响.

科学领域:

生物物理学的生物物理.
细胞生物学细胞生物学
显微镜的使用方法

背景情况:

发芽的酵母 (Saccharomyces cerevisiae) 膜蛋白Pma1对于细胞功能至关重要.
了解Pma1的扩散行为是阐明它在膜动态中的作用的关键.
光标签技术对于跟踪单颗粒水平上的蛋白质运动至关重要.

研究的目的:

调查不同的光标签策略如何影响Pma1.1测量的扩散行为.
为了比较直接融合标签的有效性与mEos3.2与新的标签结合方法.
为了验证扰动预期最大化 (pEMv2) 算法的性能,分析单粒子跟踪数据.

主要方法:

基于视频显微镜的单颗粒追踪 (SPT) 的光标记Pma1.1.
使用两种标签方法:直接融合到mEos3.2和一种新的标签绑定方案.
应用扰动预期最大化 (pEMv2) 算法来将轨迹分类为扩散状态.

主要成果:

不同的标记方法产生了显著不同的Pma1扩散分布.
对于这两种标记方法,pEMv2 确定了两种扩散状态 (不移动和移动).
与标签结合方法相比,mEos3.2的直接融合导致了更小的移动分数和更低的扩散率.
实验结果与高斯随机过程的理论预测有很好的一致性.

结论:

光标签方法的选择极大地影响了Pma1.1观察到的扩散行为.
这种新的标签绑定方法为研究Pma1动态提供了一种不那么扰乱的方法.
pEMv2算法是一种强大的工具,用于分析SPT数据和解决不同的扩散状态.

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